Media, kits, systems and methods for the micropropagation of bamboo

ABSTRACT

Disclosed herein are media, kits, systems and methods for achieving micropropagation of bamboo on a commercially-relevant scale.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority to U.S. Provisional Patent ApplicationNo. 61/304,681 filed Feb. 15, 2010 the entire disclosure of which isincorporated by reference herein.

FIELD OF THE DISCLOSURE

Disclosed herein are media, kits, systems and methods for themicropropagation of bamboo.

BACKGROUND OF THE DISCLOSURE

The subfamily Bambusoideae (of the family Poaceae), comprises both woodyand herbaceous bamboos. At present roughly 120 genera of temperate andtropical woody bamboos are recognized. Bamboos are versatile plants withmany different applications. It has been estimated that approximately2.2 billion people worldwide use bamboo to some extent, and in 1985 theglobal revenue attributable to bamboo was estimated around U.S. $4.5billion. The market for bamboo is also expanding. Bamboo shoots are astaple of Asian cuisine, and bamboo is found in a number of productsincluding toothpicks, brooms, poles for viticulture and arboriculture,landscaping materials, parquet flooring, laminate materials, furniture,handicrafts and other household items. In addition, bamboo is becomingan important source of textile material as a component of paperproduction and as a source of structural timber.

Bamboo is considered an environmentally friendly “green” product. One ofthe characteristics that gives bamboo its green reputation is itsextremely rapid growth rate. Bamboo is the fastest growing woody plantin the world, achieving growth rates of well over three feet per day. Itachieves this rate of growth in part because of its rhizome system,which is capable of providing a great deal of energy toward shootgrowth.

Despite bamboo's rapid growth rate, it has other characteristics thatmake it a difficult crop to manage. Perhaps the greatest difficultycomes from the fact that many commercially important bamboos only flowerat intervals of as long as 60-130 years. Compounding the difficulties ofthis long flowering cycle is the fact that many bamboos exhibit mass (orgregarious) flowering, with all plants in the population floweringsimultaneously. For example, Phyllostachys bambusoides flowers at aninterval of 130 years, and in this species all plants of the same stockflower at the same time, regardless of differences in geographiclocations or climatic conditions. After flowering, the bamboo dies.

Bamboo's lengthy flowering interval and propensity for mass floweringmakes it very difficult to obtain seeds for propagation. Compoundingthis problem is the fact that bamboo seeds, even when they areavailable, remain viable for no more than 3-6 months.

As a result of these difficulties with the propagation of bamboo byseed, bamboo typically is propagated by asexual techniques such as clumpdivision and cutting. These asexual propagation techniques, however, areinsufficient to meet projected world demand because both their capacityto produce mass scale production, and their practical efficiency, aretoo low. In addition many asexual propagation methods have the downsideof failing to eliminate pathogens present in the parent plants.

A method to achieve large scale production of bamboo is highlydesirable. Micropropagation (also known as tissue culturing with theterms used interchangeably herein), is an excellent method to achievethis aim.

Micropropagation is not unlike growing plants from cuttings. However,unlike plants grown from cuttings, micropropagated plants are grown invitro in sterile media. Typically, the media comprises agar, with theaddition of various compounds such as nutrients, inorganic salts, growthregulators, sugars, vitamins and other compounds.

A benefit to tissue culturing plants is that the plants can be grown ina sterile environment so that they remain disease free. Other benefitsinclude the ability to grow very large numbers of plants in a smallspace, the reduced water and nutrient needs of micropropagated plants,and the rapid multiplication of tissues that can in turn be used toyield more tissue culture material. Moreover micropropagation is veryflexible and rapid upscaling is possible (within 1 year nearly onemillion plants can be produced from any genotype). Such short timeframes and large numbers cannot be rivaled by any conventional method.Tissue culturing also provides for the production of high quality plantswhich are easy to transport and deliver.

Some papers have been published which address tissue culturing ofbamboo. In practice, however (i.e., for large or mass scale propagationof bamboos), the methods described in these papers do not translate intocommercially viable propagation systems.

The difficulties encountered in tissue culturing bamboo are highincidences of endogenous or surface contaminations and browning, factorsrelated to dormancy or topophysis and hyperhydricity. The presentdisclosure provides media, systems and methods that overcome thesedifficulties allowing the commercial-scale asexual production of bamboo.

SUMMARY OF THE DISCLOSURE

The present disclosure overcomes the difficulties encountered in thecommercial-scale asexual production of bamboo by providing effectivemedia, systems and methods for tissue culturing bamboo.

One embodiment disclosed herein is a media for micropropagating bamboowherein said media comprises meta-topolin or thidiazuron. In anotherembodiment, the media comprises meta-topolin and thidiazuron. In anotherembodiment, the media is b-9-i media, b-9-ii media, b-9-iii media,b-9-iv media, b-9-v media, CW2-i media, CW2-ii media, CW2-iii media,CW2-iv media, CW2-v media, b-10-i media, b-10-ii media, b-10-iii media,b-10-iv media, b-10-v media, b-11-i media, b-11-ii media, b-11-iiimedia, b-11-iv media, b-11-v media, b-12c-i media, b-12c-ii media,b-12c-iii media, b-12c-iv media, b-12c-v media, b-1-i media, b-1-iimedia, b-1-iii media, b-1-iv media, b-1-v media, b-4-i media, b-4-iimedia, b-4-iii media, b-4-iv media, b-4-v media, b-6-i media, b-6-iimedia, b-6-iii media, b-6-iv media, b-6-v media, CW1-i media, CW1-iimedia, CW1-iii media, CW1-iv media, CW1-v media, CW3-i media, CW3-iimedia, CW3-iii media, CW3-iv media, CW3-v media, CW4-i media, CW4-iimedia, CW4-iii media, CW4-iv media, CW4-v media, CW5-i media, CW5-iimedia, CW5-iii media, CW5-iv media, CW5-v media, CW6-i media, CW6-iimedia, CW6-iii media, CW6-iv media and/or CW6-v media.

Embodiments disclosed herein also include systems for themicropropagation of bamboo. In one embodiment, the system comprises akit comprising a media comprising meta-topolin or thidiazuron. Inanother embodiment, meta-topolin and thidiazuron are found in the samemedia or in separate media. In another embodiment, the media is b-9-imedia, b-9-ii media, b-9-iii media, b-9-iv media, b-9-v media, CW2-imedia, CW2-ii media, CW2-iii media, CW2-iv media, CW2-v media, b-10-imedia, b-10-ii media, b-10-iii media, b-10-iv media, b-10-v media,b-11-i media, b-11-ii media, b-11-iii media, b-11-iv media, b-11-vmedia, b-12c-i media, b-12c-ii media, b-12c-iii media, b-12c-iv media,b-12c-v media, b-1-i media, b-1-ii media, b-1-iii media, b-1-iv media,b-1-v media, b-4-i media, b-4-ii media, b-4-iii media, b-4-iv media,b-4-v media, b-6-i media, b-6-ii media, b-6-iii media, b-6-iv media,b-6-v media, CW1-i media, CW1-ii media, CW1-iii media, CW1-iv media,CW1-v media, CW3-i media, CW3-ii media, CW3-iii media, CW3-iv media,CW3-v media, CW4-i media, CW4-ii media, CW4-iii media, CW4-iv media,CW4-v media, CW5-i media, CW5-ii media, CW5-iii media, CW5-iv media,CW5-v media, CW6-i media, CW6-ii media, CW6-iii media, CW6-iv mediaand/or CW6-v media.

Embodiments disclosed herein also include methods of micropropagatingbamboo. In one embodiment, the method comprises culturing bambooexplants, cultures and/or shoots in media comprising meta-topolin and/orthidiazuron. In another embodiment, the meta-topolin and thidiazuron arefound in the same media or in separate media. In another embodiment, themedia is b-9-i media, b-9-ii media, b-9-iii media, b-9-iv media, b-9-vmedia, CW2-i media, CW2-ii media, CW2-iii media, CW2-iv media, CW2-vmedia, b-10-i media, b-10-ii media, b-10-iii media, b-10-iv media,b-10-v media, b-11-i media, b-11-ii media, b-11-iii media, b-11-ivmedia, b-11-v media, b-12c-i media, b-12c-ii media, b-12c-iii media,b-12c-iv media, b-12c-v media, b-1-i media, b-1-ii media, b-1-iii media,b-1-iv media, b-1-v media, b-4-i media, b-4-ii media, b-4-iii media,b-4-iv media, b-4-v media, b-6-i media, b-6-ii media, b-6-iii media,b-6-iv media, b-6-v media, CW1-i media, CW1-ii media, CW1-iii media,CW1-iv media, CW1-v media, CW3-i media, CW3-ii media, CW3-iii media,CW3-iv media, CW3-v media, CW4-i media, CW4-ii media, CW4-iii media,CW4-iv media, CW4-v media, CW5-i media, CW5-ii media, CW5-iii media,CW5-iv media, CW5-v media, CW6-i media, CW6-ii media, CW6-iii media,CW6-iv media and/or CW6-v media.

Embodiments include, without limitation:

1. A media comprising, consisting essentially of or consisting of:

mg/L (unless mg/L (unless Component otherwise noted) Component otherwisenoted) NH₄NO₃ 1650 ± 2  KH₂PO₄ 170 ± 2  KNO₃ 1900 ± 2  FeSO₄ 55.7 ± 0.2Ca(NO₃)₂ 550 ± 2  Na₂EDTA 74.6 ± 0.2 MgSO₄ 370 ± 2  Na₂H₂PO₄ 170 ± 2 MDSO₄ 16.9 ± 0.2 myo-Inositol 100 ± 2  ZnSO₄  8.6 ± 0.2 Thiamine  0.4 ±0.2 CuSO₄ 0.025 ± .002 NAA 0.05 ± .02 CaCl₂ 440 ± 2  BAP   1 ± 0.2 KI0.83 ± .02 Meta-topolin  5 ± 2 CoCl₂ 0.025 ± .002 Sugar g/L 30 ± 2 H₃BO₃ 6.2 ± .02 Agar g/L  5.5 ± 0.2 Na₂MoO₄ 0.25 ± .02 or NH₄NO₃ 1650 ± 2 KH₂PO₄ 170 ± 2  KNO₃ 1900 ± 2  FeSO₄ 55.7 ± 0.2 Ca(NO₃)₂ 550 ± 2 Na₂EDTA 74.6 ± 0.2 MgSO₄ 370 ± 2  Na₂H₂PO₄ 170 ± 2  MnSO₄ 16.9 ± 2  myo-Inositol 100 ± 2  ZnSO₄  8.6 ± 0.2 Thiamine  0.4 ± 0.2 CuSO₄ 0.025 ±.002 NAA 0.05 ± .02 CaCl₂ 440 ± 2  BAP   1 ± 0.2 KI 0.83 ± .02Thidiazuron 0.75 ± .02 CoCl₂ 0.025 ± .002 Meta-topolin  5 ± 2 H₃BO₃  6.2± 0.2 Sugar g/L 30 ± 2 Na₂MoO₄ 0.25 ± .02 Agar g/L  5.5 ± 0.22. A method of micropropagating bamboo comprising exposing a bambooexplant to a media of embodiment 1.3. A method according to embodiment 2 further comprising exposing abamboo shoot to a b-9-i media, b-9-ii media, b-9-iii media, b-9-ivmedia, b-9-v media, CW2-i media, CW2-ii media, CW2-iii media, CW2-ivmedia, CW2-v media, b-10-i media, b-10-ii media, b-10-iii media, b-10-ivmedia, b-10-v media, b-11-i media, b-11-ii media, b-11-iii media,b-11-iv media, b-11-v media, b-12c-i media, b-12c-ii media, b-12c-iiimedia, b-12c-iv media, b-12c-v media, b-1-i media, b-1-ii media, b-1-iiimedia, b-1-iv media, b-1-v media, b-4-i media, b-4-ii media, b-4-iiimedia, b-4-iv media, b-4-v media, b-6-i media, b-6-ii media, b-6-iiimedia, b-6-iv media, b-6-v media, CW1-i media, CW1-ii media, CW1-iiimedia, CW1-iv media, CW1-v media, CW3-i media, CW3-ii media, CW3-iiimedia, CW3-iv media, CW3-v media, CW4-i media, CW4-ii media, CW4-iiimedia, CW4-iv media, CW4-v media, CW5-i media, CW5-ii media, CW5-iiimedia, CW5-iv media, CW5-v media, CW6-i media, CW6-ii media, CW6-iiimedia, CW6-iv media and/or CW6-v media.4. A method according to embodiment 3 further comprising exposing abamboo shoot to a media that supports transition to ex vitro conditions.5. A method according to embodiment 3 wherein said method produces100,000 bamboo shoots from an explant.6. A method according to any of embodiments 2, 3, 4 or 5 wherein saidmethod further comprises obtaining an explant.7. A method according to embodiment 6 wherein said explant is the thirdnode from the base of a bamboo cane.8. A kit comprising a media consisting of:

mg/L (unless mg/L (unless Component otherwise noted) Component otherwisenoted) NH₄NO₃ 1650 ± 2  KH₂PO₄ 170 ± 2  KNO₃ 1900 ± 2  FeSO₄ 55.7 ± 0.2Ca(NO₃)₂ 550 ± 2  Na₂EDTA 74.6 ± 0.2 MgSO₄ 370 ± 2  Na₂H₂PO₄ 170 ± 2 MnSO₄ 16.9 ± 0.2 myo-Inositol 100 ± 2  ZnSO₄  8.6 ± 0.2 Thiamine  0.4 ±0.2 CuSO₄ 0.025 ± .002 NAA 0.05 ± .02 CaCl₂ 440 ± 2  BAP   1 ± 0.2 KI0.83 ± .02 Meta-topolin  5 ± 2 CoCl₂ 0.025 ± .002 Sugar g/L 30 ± 2 H₃BO₃ 6.2 ± .02 Agar g/L 5.5 ± 2  Na₂MoO₄ 0.25 ± .02 or NH₄NO₃ 1650 ± 2 KH₂PO₄ 170 ± 2  KNO₃ 1900 ± 2  FeSO₄ 55.7 ± 0.2 Ca(NO₃)₂ 550 ± 2 Na₂EDTA 74.6 ± 0.2 MgSO₄ 370 ± 2  Na₂H₂PO₄ 170 ± 2  MnSO₄ 16.9 ± 2  myo-Inositol 100 ± 2  ZnSO₄  8.6 ± 0.2 Thiamine  0.4 ± 0.2 CuSO₄ 0.025 ±.002 NAA 0.05 ± .02 CaCl₂ 440 ± 2  BAP   1 ± 0.2 KI 0.83 ± .02Thidiazuron 0.75 ± .02 CoCl₂ 0.025 ± .002 Meta-topolin  5 ± 2 H₃BO₃  6.2± 0.2 Sugar g/L 30 ± 2 Na₂MoO₄ 0.25 ± .02 Agar g/L  5.5 ± 0.29. A kit according to embodiment 8 further comprising a b-9-i media,b-9-ii media, b-9-iii media, b-9-iv media, b-9-v media, CW2-i media,CW2-ii media, CW2-iii media, CW2-iv media, CW2-v media, b-10-i media,b-10-ii media, b-10-iii media, b-10-iv media, b-10-v media, b-11-imedia, b-11-ii media, b-11-iii media, b-11-iv media, b-11-v media,b-12c-i media, b-12c-ii media, b-12c-iii media, b-12c-iv media, b-12c-vmedia, b-1-i media, b-1-ii media, b-1-iii media, b-1-iv media, b-1-vmedia, b-4-i media, b-4-ii media, b-4-iii media, b-4-iv media, b-4-vmedia, b-6-i media, b-6-ii media, b-6-iii media, b-6-iv media, b-6-vmedia, CW1-i media, CW1-ii media, CW1-iii media, CW1-iv media, CW1-vmedia, CW3-i media, CW3-ii media, CW3-iii media, CW3-iv media, CW3-vmedia, CW4-i media, CW4-ii media, CW4-iii media, CW4-iv media, CW4-vmedia, CW5-i media, CW5-ii media, CW5-iii media, CW5-iv media, CW5-vmedia, CW6-i media, CW6-ii media, CW6-iii media, CW6-iv media and/orCW6-v media.10. A kit according to embodiment 8 or 9 further comprising a media thatsupports transition to ex vitro conditions.11. A kit according to embodiment 8, 9, or 10 further comprising anexplant.12. A method of micropropagating bamboo comprising exposing a bambooexplant to a media comprising meta-topolin and/or thidiazuron.13. A method according to embodiment 12 wherein said media comprisesb-9-i media, b-9-ii media, b-9-iii media, b-9-iv media, b-9-v media,CW2-i media, CW2-ii media, CW2-iii media, CW2-iv media, CW2-v media,b-10-i media, b-10-ii media, b-10-iii media, b-10-iv media, b-10-vmedia, b-11-i media, b-11-ii media, b-11-iii media, b-11-iv media,b-11-v media, b-12c-i media, b-12c-ii media, b-12c-iii media, b-12c-ivmedia, b-12c-v media, b-1-i media, b-1-ii media, b-1-iii media, b-1-ivmedia, b-1-v media, b-4-i media, b-4-ii media, b-4-iii media, b-4-ivmedia, b-4-v media, b-6-i media, b-6-ii media, b-6-iii media, b-6-ivmedia, b-6-v media, CW1-i media, CW1-ii media, CW1-iii media, CW1-ivmedia, CW1-v media, CW3-i media, CW3-ii media, CW3-iii media, CW3-ivmedia, CW3-v media, CW4-i media, CW4-ii media, CW4-iii media, CW4-ivmedia, CW4-v media, CW5-i media, CW5-ii media, CW5-iii media, CW5-ivmedia, CW5-v media, CW6-i media, CW6-ii media, CW6-iii media, CW6-ivmedia and/or CW6-v media.14. A method according to embodiment 12 wherein the explant is the thirdnode from the base of a bamboo cane.15. A media comprising a b-9-i media, b-9-ii media, b-9-iii media,b-9-iv media, b-9-v media, CW2-i media, CW2-ii media, CW2-iii media,CW2-iv media, CW2-v media, b-10-i media, b-10-ii media, b-10-iii media,b-10-iv media, b-10-v media, b-11-i media, b-11-ii media, b-11-iiimedia, b-11-iv media, b-11-v media, b-12c-i media, b-12c-ii media,b-12c-iii media, b-12c-iv media, b-12c-v media, b-1-i media, b-1-iimedia, b-1-iii media, b-1-iv media, b-1-v media, b-4-i media, b-4-iimedia, b-4-iii media, b-4-iv media, b-4-v media, b-6-i media, b-6-iimedia, b-6-iii media, b-6-iv media, b-6-v media, CW1-i media, CW1-iimedia, CW1-iii media, CW1-iv media, CW1-v media, CW3-i media, CW3-iimedia, CW3-iii media, CW3-iv media, CW3-v media, CW4-i media, CW4-iimedia, CW4-iii media, CW4-iv media, CW4-v media, CW5-i media, CW5-iimedia, CW5-iii media, CW5-iv media, CW5-v media, CW6-i media, CW6-iimedia, CW6-iii media, CW6-iv media and/or CW6-v media.16. A media consisting essentially of a b-9-i media, b-9-ii media,b-9-iii media, b-9-iv media, b-9-v media, CW2-i media, CW2-ii media,CW2-iii media, CW2-iv media, CW2-v media, b-10-i media, b-10-ii media,b-10-iii media, b-10-iv media, b-10-v media, b-11-i media, b-11-iimedia, b-11-iii media, b-11-iv media, b-11-v media, b-12c-i media,b-12c-ii media, b-12c-iii media, b-12c-iv media, b-12c-v media, b-1-imedia, b-1-ii media, b-1-iii media, b-1-iv media, b-1-v media, b-4-imedia, b-4-ii media, b-4-iii media, b-4-iv media, b-4-v media, b-6-imedia, b-6-ii media, b-6-iii media, b-6-iv media, b-6-v media, CW1-imedia, CW1-ii media, CW1-iii media, CW1-iv media, CW1-v media, CW3-imedia, CW3-ii media, CW3-iii media, CW3-iv media, CW3-v media, CW4-imedia, CW4-ii media, CW4-iii media, CW4-iv media, CW4-v media, CW5-imedia, CW5-ii media, CW5-iii media, CW5-iv media, CW5-v media, CW6-imedia, CW6-ii media, CW6-iii media, CW6-iv media and/or CW6-v media.17. A media consisting of a b-9-i media, b-9-ii media, b-9-iii media,b-9-iv media, b-9-v media, CW2-i media, CW2-ii media, CW2-iii media,CW2-iv media, CW2-v media, b-10-i media, b-10-ii media, b-10-iii media,b-10-iv media, b-10-v media, b-11-i media, b-11-ii media, b-11-iiimedia, b-11-iv media, b-11-v media, b-12c-i media, b-12c-ii media,b-12c-iii media, b-12c-iv media, b-12c-v media, b-1-i media, b-1-iimedia, b-1-iii media, b-1-iv media, b-1-v media, b-4-i media, b-4-iimedia, b-4-iii media, b-4-iv media, b-4-v media, b-6-i media, b-6-iimedia, b-6-iii media, b-6-iv media, b-6-v media, CW1-i media, CW1-iimedia, CW1-iii media, CW1-iv media, CW1-v media, CW3-i media, CW3-iimedia, CW3-iii media, CW3-iv media, CW3-v media, CW4-i media, CW4-iimedia, CW4-iii media, CW4-iv media, CW4-v media, CW5-i media, CW5-iimedia, CW5-iii media, CW5-iv media, CW5-v media, CW6-i media, CW6-iimedia, CW6-iii media, CW6-iv media and/or CW6-v media.18. A kit comprising a media according to embodiment 15, 16 or 17.19. A method of micropropagating bamboo utilizing a media according toembodiment 1, 15, 16 or 17.20. A method of micropropagating bamboo utilizing a kit according toembodiment 8, 9, 10, 11 or 18.

Additional embodiments include:

21. A media comprising, consisting essentially of or consisting of:

mg/L (unless mg/L (unless Component otherwise noted) Component otherwisenoted) NH₄NO₃ 1650 ± 2  KH₂PO₄ 170 ± 2  KNO₃ 1900 ± 2  FeSO₄ 55.7 ± 0.2Ca(NO₃)₂ 550 ± 2  Na₂EDTA 74.6 ± 0.2 MgSO₄ 370 ± 2  Na₂H₂PO₄ 170 ± 2 MnSO₄ 16.9 ± 0.2 myo-Inositol 100 ± 2  ZnSO₄  8.6 ± 0.2 Thiamine  0.4 ±0.2 CuSO₄ 0.025 ± .002 NAA 0.05 ± .02 CaCl₂ 440 ± 2  BAP   1 ± 0.2 KI0.83 ± .02 Cytokinin A  5 ± 2 CoCl₂ 0.025 ± .002 Sugar g/L 30 ± 2 H₃BO₃ 6.2 ± .02 Agar g/L  5.5 ± 0.2 Na₂MoO₄ 0.25 ± .02 or NH₄NO₃ 1650 ± 2 KH₂PO₄ 170 ± 2  KNO₃ 1900 ± 2  FeSO₄ 55.7 ± 0.2 Ca(NO₃)₂ 550 ± 2 Na₂EDTA 74.6 ± 0.2 MgSO₄ 370 ± 2  Na₂H₂PO₄ 170 ± 2  MnSO₄ 16.9 ± 2  myo-Inositol 100 ± 2  ZnSO₄  8.6 ± 0.2 Thiamine  0.4 ± 0.2 CuSO₄ 0.025 ±.002 NAA 0.05 ± .02 CaCl₂ 440 ± 2  BAP   1 ± 0.2 KI 0.83 ± .02 CytokininB 0.75 ± .02 CoCl₂ 0.025 ± .002 Cytokinin A  5 ± 2 H₃BO₃  6.2 ± 0.2Sugar g/L 30 ± 2 Na₂MoO₄ 0.25 ± .02 Agar g/L  5.5 ± 0.2wherein cytokinin A is meta-topolin or a meta-topolin analogue asdescribed herein and cytokinin B is thidiazuron or a thidiazuronanalogue as described herein.22. A method of micropropagating bamboo comprising exposing a bambooexplant to a media of embodiment 21.23. A method according to embodiment 22 further comprising exposing abamboo shoot to a b-9-i media, b-9-ii media, b-9-iii media, b-9-ivmedia, b-9-v media, CW2-i media, CW2-ii media, CW2-iii media, CW2-ivmedia, CW2-v media, b-10-i media, b-10-ii media, b-10-iii media, b-10-ivmedia, b-10-v media, b-11-i media, b-11-ii media, b-11-iii media,b-11-iv media, b-11-v media, b-12c-i media, b-12c-ii media, b-12c-iiimedia, b-12c-iv media, b-12c-v media, b-1-i media, b-1-ii media, b-1-iiimedia, b-1-iv media, b-1-v media, b-4-i media, b-4-ii media, b-4-iiimedia, b-4-iv media, b-4-v media, b-6-i media, b-6-ii media, b-6-iiimedia, b-6-iv media, b-6-v media, CW1-i media, CW1-ii media, CW1-iiimedia, CW1-iv media, CW1-v media, CW3-i media, CW3-ii media, CW34 media,CW3-iv media, CW3-v media, CW4-i media, CW4-ii media, CW4-iii media,CW4-iv media, CW4-v media, CW5-i media, CW5-ii media, CW5-iii media,CW5-iv media, CW5-v media, CW6-i media, CW6-ii media, CW6-iii media,CW6-iv media and/or CW6-v media.24. A method according to embodiment 23 further comprising exposing abamboo shoot to a media that supports transition to ex vitro conditions.25. A method according to embodiment 23 wherein said method produces100,000 bamboo shoots from an explant.26. A method according to any of embodiments 22, 23, 24 or 25 whereinsaid method further comprises obtaining an explant.27. A method according to embodiment 26 wherein said explant is thethird node from the base of a bamboo cane.28. A kit comprising a media comprising, consisting essentially of orconsisting of:

mg/L (unless mg/L (unless Component otherwise noted) Component otherwisenoted) NH₄NO₃ 1650 ± 2  KH₂PO₄ 170 ± 2  KNO₃ 1900 ± 2  FeSO₄ 55.7 ± 0.2Ca(NO₃)₂ 550 ± 2  Na₂EDTA 74.6 ± 0.2 MgSO₄ 370 ± 2  Na₂H₂PO₄ 170 ± 2 MnSO₄ 16.9 ± 0.2 myo-Inositol 100 ± 2  ZnSO₄  8.6 ± 0.2 Thiamine  0.4 ±0.2 CuSO₄ 0.025 ± .002 NAA 0.05 ± .02 CaCl₂ 440 ± 2  BAP   1 ± 0.2 KI0.83 ± .02 Cytokinin A  5 ± 2 CoCl₂ 0.025 ± .002 Sugar g/L 30 ± 2 H₃BO₃ 6.2 ± .02 Agar g/L 5.5 ± 2  Na₂MoO₄ 0.25 ± .02 or NH₄NO₃ 1650 ± 2 KH₂PO₄ 170 ± 2  KNO₃ 1900 ± 2  FeSO₄ 55.7 ± 0.2 Ca(NO₃)₂ 550 ± 2 Na₂EDTA 74.6 ± 0.2 MgSO₄ 370 ± 2  Na₂H₂PO₄ 170 ± 2  MnSO₄ 16.9 ± 2  myo-Inositol 100 ± 2  ZnSO₄  8.6 ± 0.2 Thiamine  0.4 ± 0.2 CuSO₄ 0.025 ±.002 NAA 0.05 ± .02 CaCl₂ 440 ± 2  BAP   1 ± 0.2 KI 0.83 ± .02 CytokininB 0.75 ± .02 CoCl₂ 0.025 ± .002 Cytokinin A  5 ± 2 H₃BO₃  6.2 ± 0.2Sugar g/L 30 ± 2 Na₂MoO₄ 0.25 ± .02 Agar g/L  5.5 ± 0.2wherein cytokinin A is meta-topolin or a meta-topolin analogue asdescribed herein and cytokinin B is thidiazuron or a thidiazuronanalogue as described herein.29. A kit according to embodiment 28 further comprising a b-9-i media,b-9-ii media, b-9-iii media, b-9-iv media, b-9-v media, CW2-i media,CW2-ii media, CW2-iii media, CW2-iv media, CW2-v media, b-10-i media,b-10-ii media, b-10-iii media, b-10-iv media, b-10-v media, b-11-imedia, b-11-ii media, b-11-iii media, b-11-iv media, b-11-v media,b-12c-i media, b-12c-ii media, b-12c-iii media, b-12c-iv media, b-12c-vmedia, b-1-i media, b-1-ii media, b-1-iii media, b-1-iv media, b-1-vmedia, b-4-i media, b-4-ii media, b-4-iii media, b-4-iv media, b-4-vmedia, b-6-i media, b-6-ii media, b-6-iii media, b-6-iv media, b-6-vmedia, CW1-i media, CW1-ii media, CW1-iii media, CW1-iv media, CW1-vmedia, CW3-i media, CW3-ii media, CW3-iii media, CW3-iv media, CW3-vmedia, CW4-i media, CW4-ii media, CW4-iii media, CW4-iv media, CW4-vmedia, CW5-i media, CW5-ii media, CW5-iii media, CW5-iv media, CW5-vmedia, CW6-i media, CW6-ii media, CW6-iii media, CW6-iv media and/orCW6-v media.30. A kit according to embodiment 28 or 29 further comprising a mediathat supports transition to ex vitro conditions.31. A kit according to embodiment 28, 29, or 30 further comprising anexplant.32. A method of micropropagating bamboo comprising exposing a bambooexplant to a media comprising meta-topolin or a meta-topolin analogue asdescribed herein and/or thidiazuron or a thidiazuron analogue asdescribed herein.33. A method according to embodiment 32 wherein said media comprises ab-9-i media, b-9-ii media, b-9-iii media, b-9-iv media, b-9-v media,CW2-i media, CW2-ii media, CW2-iii media, CW2-iv media, CW2-v media,b-10-i media, b-10-ii media, b-10-iii media, b-10-iv media, b-10-vmedia, b-11-i media, b-11-ii media, b-11-iii media, b-11-iv media,b-11-v media, b-12c-i media, b-12c-ii media, b-12c-iii media, b-12c-ivmedia, b-12c-v media, b-1-i media, b-1-ii media, b-1-iii media, b-1-ivmedia, b-1-v media, b-4-i media, b-4-ii media, b-4-iii media, b-4-ivmedia, b-4-v media, b-6-i media, b-6-ii media, b-6-iii media, b-6-ivmedia, b-6-v media, CW1-i media, CW1-ii media, CW1-iii media, CW1-ivmedia, CW1-v media, CW3-i media, CW3-ii media, CW3-iii media, CW3-ivmedia, CW3-v media, CW4-i media, CW4-ii media, CW4-iii media, CW4-ivmedia, CW4-v media, CW5-i media, CW5-ii media, CW5-iii media, CW5-ivmedia, CW5-v media, CW6-i media, CW6-ii media, CW6-iii media, CW6-ivmedia and/or CW6-v media.34. A method according to embodiment 32 wherein the explant is the thirdnode from the base of a bamboo cane.35. A media comprising a b-9-i media, b-9-ii media, b-9-iii media,b-9-iv media, b-9-v media, CW2-i media, CW2-ii media, CW2-iii media,CW2-iv media, CW2-v media, b-10-i media, b-10-ii media, b-10-iii media,b-10-iv media, b-10-v media, b-11-i media, b-11-ii media, b-11-iiimedia, b-11-iv media, b-11-v media, b-12c-i media, b-12c-ii media,b-12c-iii media, b-12c-iv media, b-12c-v media, b-1-i media, b-1-iimedia, b-1-iii media, b-1-iv media, b-1-v media, b-4-i media, b-4-iimedia, b-4-iii media, b-4-iv media, b-4-v media, b-6-i media, b-6-iimedia, b-6-iii media, b-6-iv media, b-6-v media, CW1-i media, CW1-iimedia, CW1-iii media, CW1-iv media, CW1-v media, CW3-i media, CW3-iimedia, CW3-iii media, CW3-iv media, CW3-v media, CW4-i media, CW4-iimedia, CW4-iii media, CW4-iv media, CW4-v media, CW5-i media, CW5-iimedia, CW5-iii media, CW5-iv media, CW5-v media, CW6-i media, CW6-iimedia, CW6-iii media, CW6-iv media and/or CW6-v media.36. A media consisting essentially of a b-9-i media, b-9-ii media,b-9-iii media, b-9-iv media, b-9-v media, CW2-i media, CW2-ii media,CW2-iii media, CW2-iv media, CW2-v media, b-10-i media, b-10-ii media,b-10-iii media, b-10-iv media, b-10-v media, b-11-i media, b-11-iimedia, b-11-iii media, b-11-iv media, b-11-v media, b-12c-i media,b-12c-ii media, b-12c-iii media, b-12c-iv media, b-12c-v media, b-1-imedia, b-1-ii media, b-1-iii media, b-1-iv media, b-1-v media, b-4-imedia, b-4-ii media, b-4-iii media, b-4-iv media, b-4-v media, b-6-imedia, b-6-ii media, b-6-iii media, b-6-iv media, b-6-v media, CW1-imedia, CW1-ii media, CW1-iii media, CW1-iv media, CW1-v media, CW3-imedia, CW3-ii media, CW3-iii media, CW3-iv media, CW3-v media, CW4-imedia, CW4-ii media, CW4-iii media, CW4-iv media, CW4-v media, CW5-imedia, CW5-ii media, CW5-iii media, CW5-iv media, CW5-v media, CW6-imedia, CW6-ii media, CW6-iii media, CW6-iv media and/or CW6-v media.37. A media consisting of a b-9-i media, b-9-ii media, b-9-iii media,b-9-iv media, b-9-v media, CW2-i media, CW2-ii media, CW2-iii media,CW2-iv media, CW2-v media, b-10-i media, b-10-ii media, b-10-iii media,b-10-iv media, b-10-v media, b-11-i media, b-11-ii media, b-11-iiimedia, b-11-iv media, b-11-v media, b-12c-i media, b-12c-ii media,b-12c-iii media, b-12c-iv media, b-12c-v media, b-1-i media, b-1-iimedia, b-1-iii media, b-1-iv media, b-1-v media, b-4-i media, b-4-iimedia, b-4-iii media, b-4-iv media, b-4-v media, b-6-i media, b-6-iimedia, b-6-iii media, b-6-iv media, b-6-v media, CW1-i media, CW1-iimedia, CW1-iii media, CW1-iv media, CW1-v media, CW3-i media, CW3-iimedia, CW3-iii media, CW3-iv media, CW3-v media, CW4-i media, CW4-iimedia, CW4-iii media, CW4-iv media, CW4-v media, CW5-i media, CW5-iimedia, CW5-iii media, CW5-iv media, CW5-v media, CW6-i media, CW6-iimedia, CW6-iii media, CW6-iv media and/or CW6-v media.38. A kit comprising a media according to embodiment 35, 36 or 37.39. A method of micropropagating bamboo utilizing a media according toembodiment 21, 35, 36 or 37.40. A method of micropropagating bamboo utilizing a kit according toembodiment 28, 29, 30, 31 or 38.41. A media for micropropagating bamboo wherein said media supports10-120 day multiplication cycles at least 1 month, for at least 3months, for at least 6 months, for at least 9 months, for at least 12months, for at least 15 months, for at least 18 months, for at least 21months, for at least 24 months or for at least 36 months.42. A media according to embodiment 41 wherein said media comprises,consists essentially of or consists of b-9-i media, b-9-ii media,b-9-iii media, b-9-iv media, b-9-v media, CW2-i media, CW2-ii media,CW2-iii media, CW2-iv media, CW2-v media, b-10-i media, b-10-ii media,b-10-iii media, b-10-iv media, b-10-v media, b-11-i media, b-11-iimedia, b-11-iii media, b-11-iv media, b-11-v media, b-12c-i media,b-12c-ii media, b-12c-iii media, b-12c-iv media, b-12c-v media, b-1-imedia, b-1-ii media, b-1-iii media, b-1-iv media, b-1-v media, b-4-imedia, b-4-ii media, b-4-iii media, b-4-iv media, b-4-v media, b-6-imedia, b-6-ii media, b-6-iii media, b-6-iv media, b-6-v media, CW1-imedia, CW1-ii media, CW1-iii media, CW1-iv media, CW1-v media, CW3-imedia, CW3-ii media, CW3-iii media, CW3-iv media, CW3-v media, CW4-imedia, CW4-ii media, CW4-iii media, CW4-iv media, CW4-v media, CW5-imedia, CW5-ii media, CW5-iii media, CW5-iv media, CW5-v media, CW6-imedia, CW6-ii media, CW6-iii media, CW6-iv media and/or CW6-v media.

Additional embodiments include:

43. A media for micropropagating bamboo wherein said media comprisesmeta-topolin or an analogue thereof and supports 10-120 daymultiplication cycles for at least six months.44. A media according to embodiment 43 wherein said media supports10-120 day multiplication cycles for at least one year.45. A media according to embodiment 43 or 44 wherein said meta-topolinor analogue thereof is present in an amount from 0.0125 mg/mL-10 mg/mL.46. A media according to embodiment 43, 44, or 45 wherein said mediafurther comprises thidiazuron or an analogue thereof.47. A media according to embodiment 43, 44, 45, or 46 wherein said mediafurther comprises NAA, BAP, 2ip and/or IBA.48. A method of micropropagting bamboo comprising culturing bambooexplants and/or shoots in a media of embodiment 43, 44, 45, 46 or 47.49. A method of micropropagating bamboo according to embodiment 48wherein said bamboo is Phyllostachys bissetti; Fargesia denudata;Pleioblastus fortunei; Sasa Veitchii; Pleioblastus viridistriatus;Thamnocalamus crassinodus; Chusquea Culeo “Cana Prieta”; Bambusa OldHamii; Phyllostachys Moso; Phyllostachys Atrovaginata; DendrocalamusAsper; or Guadua Angustifolia.50. A media for transitioning shoots to ex vitro conditions whereinmedia comprises, consists essentially of or consists of Br-2-i media,Br-2-ii media, Br-2-iii media, Br-2-iv media, Br-2-v media, Ech-i media,Ech-ii media, Ech-iii media, Ech-iv, Ech-v media, Amel-i media, Amel-iimedia, Amel-iii media, Amel-iv media or Amel-v media.51. A media for micropropagating bamboo wherein said media comprisesthidiazuron or an analogue thereof and supports 10-120 daymultiplication cycles for at least six months.52. A media according to embodiment 51 wherein said media supports10-120 day multiplication cycles for at least one year.53. A media according to embodiment 51 or 52 wherein said thidiazuron oranalogue thereof is present in an amount from 0.0001 mg/mL-5 mg/mL.54. A media according to embodiment 51, 52, or 53 wherein said mediafurther comprises meta-topolin or an analogue thereof, NAA, BAP, 2ipand/or IBA.55. A media according to embodiment 54, wherein said meta-topolin oranalogue thereof is present in an amount from 0.0125 mg/mL-10 mg/mL.56. A media according to embodiment 43, 44, 45, 46, 47, 51, 52, 53, 54or 55 wherein said media comprises, consists essentially of or consistsof b-9-i media, b-9-ii media, b-9-iii media, b-9-iv media, b-9-v media,CW2-i media, CW2-ii media, CW2-iii media, CW2-iv media, CW2-v media,b-10-i media, b-10-ii media, b-10-iii media, b-10-iv media, b-10-vmedia, b-11-i media, b-11-ii media, b-11-iii media, b-11-iv media,b-11-v media, b-12c-i media, b-12c-ii media, b-12c-iii media, b-12c-ivmedia, b-12c-v media, b-1-i media, b-1-ii media, b-1-iii media, b-1-ivmedia, b-1-v media, b-4-i media, b-4-ii media, b-4-iii media, b-4-ivmedia, b-4-v media, b-6-i media, b-6-ii media, b-6-iii media, b-6-ivmedia, b-6-v media, CW1-i media, CW1-ii media, CW1-iii media, CW1-ivmedia, CW1-v media, CW3-i media, CW3-ii media, CW3-iii media, CW3-ivmedia, CW3-v media, CW4-i media, CW4-ii media, CW4-iii media, CW4-ivmedia, CW4-v media, CW5-i media, CW5-ii media, CW5-iii media, CW5-ivmedia, CW5-v media, CW6-i media, CW6-ii media, CW6-iii media, CW6-ivmedia and/or CW6-v media.57. A method of micropropagting bamboo comprising culturing bambooexplants and/or shoots in a media of embodiment 51, 52, 53, 54, or 55.58. A method of micropropagating bamboo according to embodiment 57wherein said bamboo is Phyllostachys bissetti; Fargesia denudata;Pleioblastus fortunei; Sasa Veitchii; Pleioblastus viridistriatus;Thamnocalamus crassinodus; Chusquea Culeo “Cana Prieta”; Bambusa OldHamii; Phyllostachys Moso; Phyllostachys Atrovaginata; DendrocalamusAsper; or Guadua Angustifolia.59. A kit comprising a media according to any one of embodiments 43, 44,45, 46, 47, 50, 51, 52, 53, 54, 55, or 56.

Embodiments also include all of the embodiments provided above whereinthe bamboo is that is micropropagated is Arundinaria gigantea; Bambusabalcoa; Bambusa vulgaris; Bambusa vulgaris ‘Vitatta’; Bambusa Oldhamii;Bambusa tulda; endrocalamus brandesii; Dendrocalamus asper;Dendrocalamus hamiltoni; Dendrocalamus giganteus; Dendrocalamusmembranaceus; Dendrocalamus strictus; Gigantochloa aspera; Gigantochloascortechini; Guadua culeata; uadua aculeata ‘Nicaragua’; Guaduaamplexifolia; Guadua angustifolia; Guadua angustofolia bi-color; Guaduapaniculata; Melocanna bambusoides; eohouzeaua dullooa (Teinostachyum);Ochlandra travancorica; Phyllostachys edulis ‘Moso’; Phyllostachysnigra; Phyllostachys nigra ‘Henon’; or Schizostachyum lumampao.

DETAILED DESCRIPTION OF THE DISCLOSURE

Bamboos are versatile plants with many different applications. They area staple of Asian cuisine and are found in a number of productsincluding toothpicks, brooms, poles for viticulture and arboriculture,landscaping materials, parquet flooring, laminate materials, furniture,handicrafts and other household items. In addition, bamboo is becomingan important source of textile material as a component of paperproduction and as a source of structural timber.

Bamboo is considered an environmentally friendly “green” product. One ofthe characteristics that gives bamboo its green reputation is itsextremely rapid growth rate. Despite bamboo's rapid growth rate,however, it has other characteristics that make it a difficult crop tomanage including its long flowering cycle and tendency to exhibit mass(or gregarious) flowering.

Embodiments disclosed herein provide for the micropropagation or tissueculturing (these terms are used interchangeably herein) of bamboo on acommercial scale.

Micropropagated plants are grown in vitro in sterile media. The sterilemedia can be liquid, semi-solid, or solid, and the physical state of themedia can be varied by the incorporation of one or more gelling agents.Any gelling agent known in the art that is suitable for use in planttissue culture media can be used. Agar is most commonly used for thispurpose. Examples of such agars include Agar Type A, E or M and Bacto™Agar. Other exemplary gelling agents include carrageenan, gellan gum(commercially available as PhytaGel™, Gelrite® and Gelzan™), alginicacid and its salts, and agarose. Blends of these agents, such as two ormore of agar, carrageenan, gellan gum, agarose and alginic acid or asalt thereof also can be used. Typically, the media comprises agar, withthe addition of various compounds such as nutrients, inorganic salts,growth regulators, sugars, vitamins and other compounds. Other mediaadditives can include, but are not limited to, amino acids,macroelements, iron, microelements, inositol and undefined mediacomponents such as casein hydrolysates or yeast extracts. For example,the media can include any combination of NH₄NO₃; KNO₃; Ca(NO₃)₂; K₂SO₄;MgSO₄; MnSO₄; ZnSO₄; CuSO₄; CaCl₂; Kl; CoCl₂; H₃BO₃; Na₂MoO₄; KH₂PO₄;FeSO₄; Na₂EDTA; Na₂H₂PO₄; myo-inositol; thiamine; pyridoxine; nicotinicacid; glycine; riboflavin; ascorbic acid; silicon standard solution;β-naphthoxyacetic acid (NAA); indole butyric acid (IBA); 3-indoleaceticacid (IAA); benzylaminopurine (BAP); 6-γ-γ-(dimethylallylamino)-purine(2-ip); sugar; agar; carrageenan and charcoal. Examples of plant growthregulators include auxins and compounds with auxin-like activity,cytokinins and compounds with cytokinin-like activity. Exemplary auxinsinclude 2,4-dichlorophenoxyacetic acid, IBA, picloram and combinationsthereof. Exemplary cytokinins, in addition to meta-topolin andthidiazuron, include adenine hemisulfate, benzyladenine,dimethylallyladenine, kinetin, zeatin and combinations thereof.Gibberelic acid also can be included in the media. A sugar can beincluded in the media and can serve as a carbon source. Such sugars areknown to those of ordinary skill in the art. Exemplary sugars includesucrose, glucose, maltose, galactose and sorbitol or combinationsthereof.

Disclosed herein are specialized media, systems and methods that allowthe successful tissue culturing of bamboo on a commercial scale. Certainmedia described herein include the cytokinins meta-topolin and/orthidiazuron. While certain embodiments utilize meta-topolin and/orthidiazuron defined as the particular compounds below, other relatedcompounds can also be successful.

Compounds useful according to the present disclosure includemeta-topolin analogues having a general formula

wherein W is an aryl or heteroaryl;R¹ is substituted or unsubstituted alkyl wherein any C in the alkyl canbe substituted with O, N or S;each R² is independently H, OH, C₁-C₆ alkyl, C₁-C₆ alkylene, C₁-C₆alkylyl, halogen, cyano, C₁-C₆ alkyloxy, aryl or heteroaryl eachoptionally substituted with a C₁-C₆ alkyl, SH, NHR³, CO₂R³ or halogen;R³ is H, OH, C₁-C₆ alkyl, C₁-C₆ alkylene, C₁-C₆ alkylyl, halogen,carboxylic group, ester group, aldehyde or cyano;r is 0 to 8.

In one embodiment, W is

wherein a dashed line represents the presence or absence of a bond;X1-X⁷ is each independently selected from C, N, O, S with the provisothat the X linking the ring to N is C.

In another embodiment, the compounds have a structure

wherein a dashed line represents the presence or absence of a bond.

In another embodiment, the compounds have a structure

wherein a dashed line represents the presence or absence of a bond;X⁸-X¹² is each independently selected from C, N, O, S;each R⁴ is independently H, OH, C₁-C₆ alkyl, C₁-C₆ alkylene, C₁-C₆alkylyl, halogen, cyano, C₁-C₆ alkyloxy, aryl or heteroaryl eachoptionally substituted with a C₁-C₆ alkyl, SH, NHR³, CO₂R³ or halogen;R³ is H, OH, C₁-C₆ alkyl, C₁-C₆ alkylene, C₁-C₆ alkylyl, halogen,carboxylic group, ester group, aldehyde or cyano;p is 0 to 5; andq is 0 to 6.

In other embodiments, the compounds have a structure

In still another embodiment, the compounds have a structure

Further still, compounds can have structures selected from

In one embodiment, R⁴ is OH.

In another embodiment, compounds have a structure selected from

In another embodiment, the compounds have a structure

wherein a dashed line represents the presence or absence of a bond.

In another embodiment, the compounds have a structure

wherein a dashed line represents the presence or absence of a bond;X⁸-X¹² is each independently selected from C, N, O, S;each R⁴ is independently H, OH, C₁-C₆ alkyl, C₁-C₆ alkylene, C₁-C₆alkylyl, halogen, cyano, C₁-C₆ alkyloxy, aryl or heteroaryl eachoptionally substituted with a C₁-C₆ alkyl, SH, NHR³, CO₂R³ or halogen;R³ is H, OH, C₁-C₆ alkyl, C₁-C₆ alkylene, C₁-C₆ alkylyl, halogen,carboxylic group, ester group, aldehyde or cyano;p is 0 to 5; andq is 0 to 6.

In other embodiments, the compounds have a structure

In still another embodiment, the compounds have a structure

In one embodiment, the compound is meta-topolin, also known as6-(3-hydroxybenzylamino)-purine, and by the abbreviation mT, having amolecular formula of C₁₂H₁₀N₅OH, a molecular weight of 241.25, and thefollowing structural formula:

wherein said meta-topolin is a derivative of a willow tree or a poplartree.

Compounds useful according to the present disclosure includethiadiazuron analogues having a general formula

wherein V is an aryl or heteroaryl;each R⁵ and R⁶ is each independently H, OH, C₁-C₆ alkyl, C₁-C₆ alkylene,C₁-C₆ alkylyl, halogen, cyano, C₁-C₆ alkyloxy, aryl or heteroaryl eachoptionally substituted with a C₁-C₆ alkyl or halogen;n is 0 to 4;o is 0 to 5X¹³-X¹⁶ is each independently selected from C, N, O, S;Z¹ and Z² are each independently NH, O, SH or CH or Z¹ and Z² can becombined to form a substituted or unsubstituted aryl or heteroaryl; and

Y¹ is O or S.

In another embodiment, compounds have a structure

wherein X¹⁷-X²¹ is each independently selected from C, N, O, S.

In other embodiments, compounds include

In one embodiment, the compound is Thidiazuron, also known as1-phenyl-3-(1,2,3-thiadiazol-5-yl)urea and5-phenylcarbamoylamino-1,2,3-thiadiazole has the molecular formula ofC₉H₈N₄OS, a molecular weight of 220.25 and the following structuralformula

If present in a media, each cytokinin can be present in an amount from0.001 mg/L-100 mg/L and all amounts in between. In certain embodiments,meta-topolin and/or its analogues can be present at 0.001 mg/L, 0.01,0.1, 1, 2, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2,4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7,5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2,7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53,54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 mg/L. In particularembodiments, thidiazuron and/or its analogues can be present at 0.001mg/L, 0.01, 0.025, 0.05, 0.075, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4,0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1.0, 1.25,1.50, 1.75, 2.25, 2.5, 2.75, 3.5, 4.5, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49,50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67,68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85,86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 mg/L.

When both are utilized, meta-topolin and/or its analogues andthidiazuron and/or its analogues can also be included in ratios. Forexample, the amount of meta-topolin and/or its analogues to thidiazuronand/or its analogues can be 100:1, 95:1, 90:1, 85:1, 80:1, 75:1, 70:1,65:1, 60:1, 55:1, 50:1, 45:1, 40:1, 35:1, 30:1, 29:1, 28:1, 27:1, 26:1,25:1, 24:1, 23:1, 22:1, 21:1, 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1,13:1, 12:1, 11:1, 10:1; 9:1, 8:1, 7:1, 6.9:1, 6.8:1, 6.7:1, 6.6:1,6.5:1, 6.4:1, 6.3:1, 6.2:1, 6.1:1, 6:1, 5.9:1, 5.8:1, 5.7:1, 5.6:1,5.5:1, 5.4:1, 5.3:1, 5.2:1, 5.1:1, 5:1; 4:1, 3:1, 2:1, 1:1, 0.75:1,0.5:1, 0.25:1, 0.1:1, 0.075:1, 0.05:1, 0.025:1 or 0.001:1. When bothmeta-topolin and thidiazuron are used, they can be present in the sameor different media.

NAA, BAP, 2ip and/or IBA can similarly can be present at 0.001 mg/L,0.01, 0.1, 1, 2, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1,4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6,5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1,7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51,52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 mg/L or 0.001mg/L, 0.01, 0.025, 0.05, 0.075, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4,0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1.0, 1.25,1.50, 1.75, 2.25, 2.5, 2.75, 3.5, 4.5, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49,50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67,68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85,86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 mg/L.

The structures or formula for a number of chemical compounds, includingmeta-topolin and thidiazuron, have been provided above. One of ordinaryskill in the art will recognize reference to a compound should beconstrued broadly to include pharmaceutically acceptable salts,prodrugs, tautomers, alternate solid forms, non-covalent complexes,analogs, derivatives and combinations thereof, of a chemical entity ofthe depicted structure or chemical name.

A pharmaceutically acceptable salt is any salt of the parent compoundthat is suitable for use in the methods disclosed herein. Apharmaceutically acceptable salt also refers to any salt which may formas a result of administration of an acid, another salt, or a prodrugwhich is converted into an acid or salt. A salt comprises one or moreionic forms of the compound, such as a conjugate acid or base,associated with one or more corresponding counter-ions. Salts can formfrom or incorporate one or more deprotonated acidic groups (e.g.carboxylic acids), one or more protonated basic groups (e.g. amines), orboth (e.g. zwitterions).

Not intended to be limited by the above described compounds, varioustautomers of the above compounds may be possible. As used herein,“tautomer” refers to the migration of protons between adjacent singleand double bonds. The tautomerization process is reversible. Othertautomers are possible when the compound includes, for example but notlimited to, enol, keto, lactamin, amide, imidic acid, amine, and iminegroups. Tautomers will generally reach an equilibrium state wherein thedouble bond is resonantly shared between the two bond lengths.

Unless stereochemistry is explicitly depicted, a structure is intendedto include every possible stereoisomer, both pure or in any possiblemixture.

Alternate solid forms are different solid forms than those that mayresult from practicing the procedures described herein. For example,alternate solid forms may be polymorphs, different kinds of amorphoussolid forms, glasses, and the like.

Non-covalent complexes are complexes that may form between the compoundand one or more additional chemical species that do not involve acovalent bonding interaction between the compound and the additionalchemical species. They may or may not have a specific ratio between thecompound and the additional chemical species. Examples might includesolvates, hydrates, charge transfer complexes, and the like.

As an overview, in typical micropropagation, plants are placed invarious media that stimulate physiological processes such as growth andmultiplication by and/or within the plant. Generally the processincludes 3 steps (following explant preparation and disinfection,discussed below): (1) initiation of in vitro growth and/ormultiplication of the explant in a media; (2) further in vitromultiplication in a second media; and (3) transition to ex vitroconditions. Not every tissue culture process requires each step,however, and in certain processes, steps can be combined or skipped. Forexample, while there is commonly a change in media types between steps 1and 2, in certain embodiments, a media change is not included. In otherprocesses, plants may not require a particular step promoting transitionto ex vitro conditions but instead complete the process in a same mediathat supports multiplication. Accordingly, as described herein, mediaare defined as Stage 1 media (1^(st) media of a process); Stage 2 media(2^(nd) media of a process); Stage 3 media (3^(rd) media of a process);etc. Particular media can change stage based on the number of stepswithin a particular process and where the particular media resideswithin their order.

To begin the process, a Stage 1 media can be obtained or prepared. Stage1 media include a pH that is generally hospitable to plants (typicallyfrom 4.0-7.0 or 4.5-6.5). The Stage 1 media is then placed into testtubes or other appropriate containers (including jars, boxes, jugs,cups, etc. wherein when not specified are collectively referred to as“tubes”). These tubes can be capped or covered and autoclaved tosterilize the tubes and media. In another embodiment, sterilization isachieved by autoclaving at 5-25 pounds pressure psi at a temperature of200° F.-for 200° F. 10-25 minutes. In another embodiment, sterilizationis achieved by autoclaving at 15 pounds pressure psi at a temperature of250° F. for 15-18 minutes. Sterility can also be assessed by an acceptednumber of contaminated tubes per hundred tubes, for example and withoutlimitation, 0 contaminated tubes per hundred tubes, no more than 1contaminated tube per hundred tubes, no more than 2 contaminated tubesper hundred tubes, no more than 3 contaminated tubes per hundred tubes,no more than 4 contaminated tubes per hundred tubes, no more than 5contaminated tubes per hundred tubes, no more than 6 contaminated tubesper hundred tubes, no more than 7 contaminated tubes per hundred tubes,no more than 8 contaminated tubes per hundred tubes, no more than 9contaminated tubes per hundred tubes, no more than 10 contaminated tubesper hundred tubes, etc.

In media containing a gelling agent, such as agar, agarose, gellan gum,carrageenan or combinations thereof, the media solidifies upon coolingand serves to provide the micropropagated plant tissues with support,nutrients, growth regulators, water and other compounds as describedbelow. Generally, tubes and jars contain 15-25 mL media while boxescontain 40-50 mL media. Cups can include 30-40 mL while jugs generallycontain more than 50 mL.

Micropropagated plants begin from a selected piece of plant tissue,called an “explant” or “mother plant.” This explant is the source ofcells to be developed during the tissue culturing process. The explantcan be any segment or collection of cells from apical meristem, axillarybuds, cambium, lateral meristem, shoot apices, stem segments, immaturenodal sections from stems, lateral shoots, seedlings or leaf segments.In one embodiment, the explant is taken from a 1 year old bamboo plant.In another embodiment, the explant is taken from a 2 year old bambooplant. In another embodiment, the explant is taken from a bamboo plantthat is 5 years old or less. In another embodiment, the explant is takenfrom a bamboo plant that is 4 years old or less. In another embodiment,the explant is taken from a bamboo plant that is 3 years old or less. Inanother embodiment, the explant is taken from a bamboo plant that is 2years old or less. In another embodiment, the explant is taken from abamboo plant that is 1 years old or less. In another embodiment, theexplant is taken from a bamboo plant that is 6 months old or less. Inanother embodiment, the explant is taken from a bamboo plant that is 3months old or less. The bamboo from which the explant is obtained can begrown in any suitable husbandry situation, including but not limited togrowing in a growth chamber, growing in a greenhouse or growing in afield.

As will be understood by one of ordinary skill in the art, a variety ofappropriate explants can be used in accordance with the presentdisclosure. In certain embodiments according to the present disclosure,immature nodal sections from stems can be used as the explant material.In one embodiment, the explants can be new growth canes with the lateralshoots just breaking the sheath at nodal section(s). New growth canesinclude those obtained from the plant within a current season or year,wherein such new growth canes can be obtained from any node on theplant. In one particular embodiment, explant material includes or islimited to the third node from the base of a cane.

Nodal section(s) can be cut into 3-5, 1-10, 2-9, 3-8, 4-6, 3-6 or 2-7millimeter sections with the shoot intact and disinfected to removepathogens on the exterior of the explant. Any disinfection method knownin the art can be used. Exemplary disinfection methods includeapplication of a disinfectant, such as a disinfectant selected fromamong bleach (sodium and/or potassium and/or calcium hypochlorite),alcohol (e.g., ethanol, isopropyl), ozone, chlorine gas, iodine solutionor antibiotic or anti-fungal solution or combinations thereof, orsubjecting the exposed surface of the explant to ultraviolet light or toa temperature of −20° C. or lower or to a temperature higher than 40° C.or 50° C. for a short period of time. In certain embodiments, smallamounts (a few drops) of Tween 20 can be added to the disinfectingsolutions.

Following initial disinfection, the outer sheaths can be peeled off anddiscarded and the remaining piece put into an approximately 1%, 5%, 10%,15%, 20%, 25% or 30% solution of a commercial bleach or a similardisinfecting solution. The peeled explant in disinfecting solution canbe put onto a shaker table, such as for example, a Lab Rotators,Adjustable speed, Barnstead/Lab line orbital Shaker (model number KS260) for 10 minutes, 20 minutes, 30 minutes, 60 minutes, 90 minutes, 120minutes, 150 minutes, 180 minutes or 210 minutes at 6-9 or 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11 or 12, 13, 14, or 15 revolutions per minute. Inanother embodiment, the peeled explants can then be put into anapproximately 1% solution of bleach or similar disinfecting solution,and placed back onto the shaker table for 30 minutes. In anotherembodiment, this 1% bleach or similar disinfecting solution step can berepeated. In another embodiment, these described steps are progressiveand include the entire disinfection process. As will be understood byone of ordinary skill in the art, a variety of appropriate disinfectingprocedures can be used in accordance with the present disclosure.

Once disinfected, the explants can be placed onto a Stage 1 media withinthe tube and the tubes can be placed in a regulated growth chamber. Asused herein, “growth chambers” can include a number of configurationsand sizes including table-top boxes, stand-alone chambers, closets,small rooms, large rooms, etc. As is understood by one of ordinary skillin the art, variables such as light or temperature can be appropriatelycontrolled in such a growth chamber. Appropriate ranges for tissueculturing bamboo include from 65° F.-70° F., 60° F.-75° F. or 55° F.-80°F. at 200-500, 150-550 or 100-600 foot candles. Lighting can be fullspectrum, although alternative lighting systems can also be utilizedaccording to the present disclosure.

The explants are allowed to establish themselves within the tubes whilein the growth chamber on Stage 1 media. In more common 3 stage tissueculturing, once established (i.e. growing without visiblecontamination), the cell cultures grown from the explants aretransferred into a second, Stage 2 media. Alternatively, onceestablished, the cell cultures can remain in Stage 1 media. At thisstage in the tissue culturing process, a large number of plants can becreated within a relatively short period of time because each cellculture can develop multiple shoots and each shoot can be separated andplaced into an individual tube where it will develop additional shootsto separate and multiply.

Without limiting the media to a particular stage, non-limiting examplesof media that commonly serve as Stage 1 and/or Stage 2 media include:

Media b-12c(i-v):

Component (mg/L in all unless otherwise noted) b-12c-i b-12c-iib-12c-iii b-12c-iv b-12c-v NH₄NO₃  825-2475 1237-2063 1485-1815 16501650 ± 2   KNO₃  950-2850 1425-2375 1710-2090 1900 1900 ± 2   Ca(NO₃)₂225-775 410-690 495-605 550 550 ± 2  MgSO₄ 185-555 275-465 330-410 370370 ± 2  MnSO₄  8.0-26.0 12.0-22.0 15.0-19.0 16.9 16.9 ± 0.2  ZnSO₄ 4.0-12.0  6.0-10.0 8.0-9.0 8.6 8.6 ± 0.2 CuSO₄ 0.012-0.378 0.020-0.0300.022-0.028 0.025 0.025 ± .002  CaCl₂ 220-660 330-350 400-480 440 440 ±2  KI 0.40-1.25 0.60-1.05 0.75-0.90 0.83 0.83 ± .02  CoCl₂ 0.012-0.3780.020-0.030 0.022-0.028 0.025 0.025 ± .002  H₃BO₃ 3.0-9.0 4.0-8.05.0-7.0 6.2 6.2 ± 0.2 Na₂MoO₄ 0.12-0.36 0.18-0.31 .22-.28 0.25 0.25 ±.02  KH₂PO₄  85-255 120-210 150-190 170 170 ± 2  FeSO₄ 27.0-84.040.0-70.0 50.0-60.0 55.7 55.7 ± 0.2  Na₂EDTA  37.0-112.0 55.0-94.067.0-82.0 74.6 74.6 ± 0.2  Na₂H₂PO₄  85-255 120-210 150-190 170 170 ± 2 myo-Inositol  50-150  75-125  90-110 100 100 ± 2  Thiamine 0.2-0.60.3-0.5 0.36-0.44 0.4 0.4 ± 0.2 NAA 0.02-0.08 0.03-0.07 0.04-0.06 0.050.05 ± .02  BAP 0.5-1.5 0.7-1.3 0.9-1.1 1   1 ± 0.2 Thidiazuron0.36-1.12 0.56-0.94 0.67-.083 0.75 0.75 ± .02  Meta-topolin 2.5-7.53.7-6.2 4.5-5.5 5 5 ± 2 Sugar g/L 15-45 22-37 27-33 30 30 ± 2  Agar g/L2.7-8.2 4.1-6.8 4.9-6.1 5.5 5.5 ± 0.2Media CW2(i-v):

Component CW2-i CW2-ii CW2-iii CW2-iv CW2-v NH₄NO₃  825-2475 1237-20631485-1815 1650 1650 ± 2   KNO₃  950-2850 1425-2375 1710-2090 1900 1900 ±2   Ca(NO₃)₂ 225-775 410-690 495-605 550 550 ± 2  MgSO₄ 185-555 275-465330-410 370 370 ± 2  MnSO₄  8.0-26.0 12.0-22.0 15.0-19.0 16.9 16.9 ±0.2  ZnSO₄  4.0-12.0  6.0-10.0 8.0-9.0 8.6 8.6 ± 0.2 CuSO₄ 0.012-0.3780.020-0.030 0.022-0.028 0.025 0.025 ± .002  CaCl₂ 220-660 330-350400-480 440 440 ± 2  KI 0.40-1.25 0.60-1.05 0.75-0.90 0.83 0.83 ± .02 CoCl₂ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ± .002  H₃BO₃3.0-9.0 4.0-8.0 5.0-7.0 6.2 6.2 ± 0.2 Na₂MoO₄ 0.12-0.36 0.18-0.31.22-.28 0.25 0.25 ± .02  KH₂PO₄  85-255 120-210 150-190 170 170 ± 2 FeSO₄ 27.0-84.0 40.0-70.0 50.0-60.0 55.7 55.7 ± 0.2  Na₂EDTA  37.0-112.055.0-94.0 67.0-82.0 74.6 74.6 ± 0.2  Na₂H₂PO₄  85-255 120-210 150-190170 170 ± 2  myo-Inositol  50-150  75-125  90-110 100 100 ± 2  Thiamine0.2-0.6 0.3-0.5 0.36-0.44 0.4 0.4 ± 0.2 NAA 0.02-0.08 0.03-0.070.04-0.06 0.05 0.05 ± .02  BAP 0.5-1.5 0.7-1.3 0.9-1.1 1   1 ± 0.2Meta-topolin 2.5-7.5 3.7-6.2 4.5-5.5 5 5 ± 2 Sugar g/L 15-45 22-37 27-3330 30 ± 2  Agar g/L 2.7-8.2 4.1-6.8 4.9-6.1 5.5 5.5 ± 0.2Media CW3(i-v):

Component CW3-i CW3-ii CW3-iii CW3-iv CW3-v NH₄NO₃  825-2475 1237-20631485-1815 1650 1650 ± 2   KNO₃  950-2850 1425-2375 1710-2090 1900 1900 ±2   Ca(NO₃)₂ 225-775 410-690 495-605 550 550 ± 2  MgSO₄ 185-555 275-465330-410 370 370 ± 2  MnSO₄  8.0-26.0 12.0-22.0 15.0-19.0 16.9 16.9 ±0.2  ZnSO₄  4.0-12.0  6.0-10.0 8.0-9.0 8.6 8.6 ± 0.2 CuSO₄ 0.012-0.3780.020-0.030 0.022-0.028 0.025 0.025 ± .002  CaCl₂ 220-660 330-350400-480 440 440 ± 2  KI 0.40-1.25 0.60-1.05 0.75-0.90 0.83 0.83 ± .02 CoCl₂ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ± .002  H₃BO₃3.0-9.0 4.0-8.0 5.0-7.0 6.2 6.2 ± 0.2 Na₂MoO₄ 0.12-0.36 0.18-0.31.22-.28 0.25 0.25 ± .02  KH₂PO₄  85-255 120-210 150-190 170 170 ± 2 FeSO₄ 27.0-84.0 40.0-70.0 50.0-60.0 55.7 55.7 ± 0.2  Na₂EDTA  37.0-112.055.0-94.0 67.0-82.0 74.6 74.6 ± 0.2  Na₂H₂PO₄  85-255 120-210 150-190170 170 ± 2  myo-Inositol  50-150  75-125  90-110 100 100 ± 2  Thiamine0.2-0.6 0.3-0.5 0.36-0.44 0.4 0.4 ± 0.2 NAA 0.05-0.15 0.07-0.120.09-0.11 0.1  0.1 ± 0.02 BAP 0.5-1.5 0.7-1.3 0.9-1.1 1   1 ± 0.2 IBA0.1-0.3 0.15-0.25 0.17-0.22 0.2 0.2 ± 0.1 Meta-topolin 2.5-7.5 3.7-6.24.5-5.5 5 5 ± 2 Sugar g/L 15-45 22-37 27-33 30 30 ± 2  Agar g/L 2.7-8.24.1-6.8 4.9-6.1 5.5 5.5 ± 0.2Media b-9(i-v):

Component b-9-i b-9-ii b-9-iii b-9-iv b-9-v NH₄NO₃  825-2475 1237-20631485-1815 1650 1650 ± 2   KNO₃  950-2850 1425-2375 1710-2090 1900 1900 ±2   MgSO₄ 185-555 275-465 330-410 370 370 ± 2  MnSO₄  8.0-26.0 12.0-22.015.0-19.0 16.9 16.9 ± 0.2  ZnSO₄  4.0-12.0  6.0-10.0 8.0-9.0 8.6 8.6 ±0.2 CuSO₄ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ± .002  CaCl₂220-660 330-350 400-480 440 440 ± 2  KI 0.40-1.25 0.60-1.05 0.75-0.900.83 0.83 ± .02  CoCl₂ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ±.002  H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2 6.2 ± 0.2 Na₂MoO₄ 0.12-0.360.18-0.31 .22-.28 0.25 0.25 ± .02  KH₂PO₄  85-255 120-210 150-190 170170 ± 2  FeSO₄ 27.0-84.0 40.0-70.0 50.0-60.0 55.7 55.7 ± 0.2  Na₂EDTA 37.0-112.0 55.0-94.0 67.0-82.0 74.6 74.6 ± 0.2  Na₂H₂PO₄  85-255120-210 150-190 170 170 ± 2  myo-Inositol  50-150  75-125  90-110 100100 ± 2  Thiamine 0.2-0.6 0.3-0.5 0.36-0.44 0.4 0.4 ± 0.2 NAA 0.02-0.080.03-0.07 0.04-0.06 0.05 0.05 ± .02  BAP 0.5-1.5 0.7-1.3 0.9-1.1 1   1 ±0.2 Thidiazuron 0.12-0.36 0.18-0.31 .22-.28 0.25 0.25 ± .02 Meta-topolin 2.5-7.5 3.7-6.2 4.5-5.5 5 5 ± 2 Sugar g/L 15-45 22-37 27-3330 30 ± 2  Agar 2.7-8.2 4.1-6.8 4.9-6.1 5.5 5.5 ± 0.2 g/LMedia CW4(i-v):

CW4- Component CW4-i CW4-ii CW4-iii iv CW4-v NH₄NO₃  825-2475 1237-20631485-1815 1650  1650 ± 2 KNO₃  950-2850 1425-2375 1710-2090 1900  1900 ±2 MgSO₄ 185-555 275-465 330-410 370   370 ± 2 MnSO₄  8.0-26.0 12.0-22.015.0-19.0 16.9  16.9 ± 0.2 ZnSO₄  4.0-12.0  6.0-10.0 8.0-9.0 8.6  8.6 ±0.2 CuSO₄ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ± .002 CaCl₂220-660 330-350 400-480 440   440 ± 2 KI 0.40-1.25 0.60-1.05 0.75-0.900.83  0.83 ± .02 CoCl₂ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ±.002 H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2  6.2 ± 0.2 Na₂MoO₄ 0.12-0.360.18-0.31 .22-.28 0.25  0.25 ± .02 KH₂PO₄  85-255 120-210 150-190 170  170 ± 2 FeSO₄ 27.0-84.0 40.0-70.0 50.0-60.0 55.7  55.7 ± 0.2 Na₂EDTA 37.0-112.0 55.0-94.0 67.0-82.0 74.6  74.6 ± 0.2 Na₂H₂PO₄  85-255120-210 150-190 170   170 ± 2 myo-Inositol  50-150  75-125  90-110 100  100 ± 2 Thiamine 0.2-0.6 0.3-0.5 0.36-0.44 0.4  0.4 ± 0.2 NAA0.05-0.15 0.07-0.12 0.09-0.11 0.1  0.1 ± 0.02 BAP 0.5-1.5 0.7-1.30.9-1.1 1    1 ± 0.2 IBA 0.1-0.3 0.15-0.25 0.17-0.22 0.2  0.2 ± 0.1Thidiazuron 0.12-0.36 0.18-0.31 .22-.28 0.25  0.25 ± .02 Meta-topolin2.5-7.5 3.7-6.2 4.5-5.5 5    5 ± 2 Sugar g/L 15-45 22-37 27-33 30   30 ±2 Agar 2.7-8.2 4.1-6.8 4.9-6.1 5.5  5.5 ± 0.2 g/LMedia CW1(i-v):

CW1- Component CW1-i CW1-ii CW1-iii iv CW1-v NH₄NO₃  825-2475 1237-20631485-1815 1650  1650 ± 2 KNO₃  950-2850 1425-2375 1710-2090 1900  1900 ±2 MgSO₄ 185-555 275-465 330-410 370   370 ± 2 MnSO₄  8.0-26.0 12.0-22.015.0-19.0 16.9  16.9 ± 0.2 ZnSO₄  4.0-12.0  6.0-10.0 8.0-9.0 8.6  8.6 ±0.2 CuSO₄ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ± .002 CaCl₂220-660 330-350 400-480 440   440 ± 2 KI 0.40-1.25 0.60-1.05 0.75-0.900.83  0.83 ± .02 CoCl₂ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ±.002 H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2  6.2 ± 0.2 Na₂MoO₄ 0.12-0.360.18-0.31 .22-.28 0.25  0.25 ± .02 KH₂PO₄  85-255 120-210 150-190 170  170 ± 2 FeSO₄ 27.0-84.0 40.0-70.0 50.0-60.0 55.7  55.7 ± 0.2 Na₂EDTA 37.0-112.0 55.0-94.0 67.0-82.0 74.6  74.6 ± 0.2 Na₂H₂PO₄  85-255120-210 150-190 170   170 ± 2 myo-Inositol  50-150  75-125  90-110 100  100 ± 2 Thiamine 0.2-0.6 0.3-0.5 0.36-0.44 0.4  0.4 ± 0.2 NAA0.02-0.08 0.03-0.07 0.04-0.06 0.05  0.05 ± .02 BAP 0.5-1.5 0.7-1.30.9-1.1 1    1 ± 0.2 Meta-topolin 2.5-7.5 3.7-6.2 4.5-5.5 5    5 ± 2Sugar g/L 15-45 22-37 27-33 30   30 ± 2 Agar g/L 2.7-8.2 4.1-6.8 4.9-6.15.5  5.5 ± 0.2 Silicon 0.5-1.5 0.7-1.3 0.9-1.1 1    1 ± 0.2 Solution mLMedia CW5(i-v):

CW5- Component CW5-i CW5-ii CW5-iii iv CW5-v NH₄NO₃  825-2475 1237-20631485-1815 1650  1650 ± 2 KNO₃  950-2850 1425-2375 1710-2090 1900  1900 ±2 MgSO₄ 185-555 275-465 330-410 370   370 ± 2 MnSO₄  8.0-26.0 12.0-22.015.0-19.0 16.9  16.9 ± 0.2 ZnSO₄  4.0-12.0 6.0-10.0 8.0-9.0 8.6  8.6 ±0.2 CuSO₄ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ± .002 CaCl₂220-660 330-350 400-480 440   440 ± 2 KI 0.40-1.25 0.60-1.05 0.75-0.900.83  0.83 ± .02 CoCl₂ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ±.002 H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2  6.2 ± 0.2 Na₂MoO₄ 0.12-0.360.18-0.31 .22-.28 0.25  0.25 ± .02 KH₂PO₄  85-255 120-210 150-190 170  170 ± 2 FeSO₄ 27.0-84.0 40.0-70.0 50.0-60.0 55.7  55.7 ± 0.2 Na₂EDTA 37.0-112.0 55.0-94.0 67.0-82.0 74.6  74.6 ± 0.2 Na₂H₂PO₄  85-255120-210 150-190 170   170 ± 2 myo-Inositol  50-150  75-125  90-110 100  100 ± 2 Thiamine 0.2-0.6 0.3-0.5 0.36-0.44 0.4  0.4 ± 0.2 NAA0.02-0.08 0.03-0.07 0.04-0.06 0.05  0.05 ± .02 BAP 0.5-1.5 0.7-1.30.9-1.1 1    1 ± 0.2 IBA 0.1-0.3 0.15-0.25 0.17-0.22 0.2  0.2 ± 0.1Meta-topolin 2.5-7.5 3.7-6.2 4.5-5.5 5    5 ± 2 Sugar g/L 15-45 22-3727-33 30   30 ± 2 Agar g/L 2.7-8.2 4.1-6.8 4.9-6.1 5.5  5.5 ± 0.2Silicon 0.5-1.5 0.7-1.3 0.9-1.1 1    1 ± 0.2 Solution mLMedia CW6(i-v):

CW6- Component CW6-i CW6-ii CW6-iii iv CW6-v NH₄NO₃  825-2475 1237-20631485-1815 1650   1650 ± 2 KNO₃  950-2850 1425-2375 1710-2090 1900   1900± 2 MgSO₄ 185-555 275-465 330-410 370   370 ± 2 MnSO₄  8.0-26.012.0-22.0 15.0-19.0 16.9  16.9 ± 0.2 ZnSO₄  4.0-12.0  6.0-10.0 8.0-9.08.6   8.6 ± 0.2 CuSO₄ 0.012-0.378 0.020-0.030 0.022-0.028 0.025  0.025 ±.002 CaCl₂ 220-660 330-350 400-480 440   440 ± 2 KI 0.40-1.25 0.60-1.050.75-0.90 0.83  0.83 ± .02 CoCl₂ 0.012-0.378 0.020-0.030 0.022-0.0280.025  0.025 ± .002 H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2   6.2 ± 0.2Na₂MoO₄ 0.12-0.36 0.18-0.31 .22-.28 0.25  0.25 ± .02 KH₂PO₄  85-255120-210 150-190 170   170 ± 2 K₂SO₅ 181.85-545.63 272.80-454.69327.45-400.05 363.75 363.75 ± .02 FeSO₄ 27.0-84.0 40.0-70.0 50.0-60.055.7  55.7 ± 0.2 Na₂EDTA  37.0-112.0 55.0-94.0 67.0-82.0 74.6  74.6 ±0.2 Na₂H₂PO₄  85-255 120-210 150-190 170   170 ± 2 myo-Inositol  50-150 75-125  90-110 100   100 ± 2 Thiamine 0.2-0.6 0.3-0.5 0.36-0.44 0.4  0.4 ± 0.2 NAA 0.05-0.15 0.07-0.12 0.09-0.11 0.1   0.1 ± 0.02 BAP0.5-1.5 0.7-1.3 0.9-1.1 1    1 ± 0.2 IBA 0.1-0.3 0.15-0.25 0.17-0.22 0.2  0.2 ± 0.1 Meta-topolin 2.5-7.5 3.7-6.2 4.5-5.5 5    5 ± 2 Thidiazuron0.12-0.36 0.18-0.31 .22-.28 0.25  0.25 ± .02 Sugar g/L 15-45 22-37 27-3330    30 ± 2 Agar g/L 2.7-8.2 4.1-6.8 4.9-6.1 5.5   5.5 ± 0.2Media b-100-v):

b-10- Component b-10-i b-10-ii b-10-iii iv b-10-v NH₄NO₃  825-24751237-2063 1485-1815 1650  1650 ± 2 KNO₃  950-2850 1425-2375 1710-20901900  1900 ± 2 MgSO₄ 185-555 275-465 330-410 370   370 ± 2 MnSO₄ 8.0-26.0 12.0-22.0 15.0-19.0 16.9  16.9 ± 0.2 ZnSO₄  4.0-12.0  6.0-10.08.0-9.0 8.6  8.6 ± 0.2 CuSO₄ 0.012-0.378 0.020-0.030 0.022-0.028 0.0250.025 ± .002 CaCl₂ 220-660 330-350 400-480 440   440 ± 2 KI 0.40-1.250.60-1.05 0.75-0.90 0.83  0.83 ± .02 CoCl₂ 0.012-0.378 0.020-0.0300.022-0.028 0.025 0.025 ± .002 H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2  6.2 ±0.2 Na₂MoO₄ 0.12-0.36 0.18-0.31 .22-.28 0.25  0.25 ± .02 KH₂PO₄  85-255120-210 150-190 170   170 ± 2 FeSO₄ 27.0-84.0 40.0-70.0 50.0-60.0 55.7 55.7 ± 0.2 Na₂EDTA  37.0-112.0 55.0-94.0 67.0-82.0 74.6  74.6 ± 0.2Na₂H₂PO₄  85-255 120-210 150-190 170   170 ± 2 myo-Inositol  50-150 75-125  90-110 100   100 ± 2 Thiamine 0.2-0.6 0.3-0.5 0.36-0.44 0.4 0.4 ± 0.2 NAA 0.02-0.08 0.03-0.07 0.04-0.06 0.05  0.05 ± .02 BAP0.5-1.5 0.7-1.3 0.9-1.1 1    1 ± 0.2 Meta-topolin 2.5-7.5 3.7-6.24.5-5.5 5    5 ± 2 Sugar g/L 15-45 22-37 27-33 30   30 ± 2 Agar g/L2.5-7.5 3.7-6.2 4.5-5.5 5    5 ± 2Media b-11(i-v):

b-11- Component b-11-i b-11-ii b-11-iii iv b-11-v NH₄NO₃  825-24751237-2063 1485-1815 1650  1650 ± 2 KNO₃  950-2850 1425-2375 1710-20901900  1900 ± 2 MgSO₄ 185-555 275-465 330-410 370   370 ± 2 MnSO₄ 8.0-26.0 12.0-22.0 15.0-19.0 16.9  16.9 ± 0.2 ZnSO₄  4.0-12.0  6.0-10.08.0-9.0 8.6  8.6 ± 0.2 CuSO₄ 0.012-0.378 0.020-0.030 0.022-0.028 0.0250.025 ± .002 CaCl₂ 220-660 330-350 400-480 440   440 ± 2 KI 0.40-1.250.60-1.05 0.75-0.90 0.83  0.83 ± .02 CoCl₂ 0.012-0.378 0.020-0.0300.022-0.028 0.025 0.025 ± .002 H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2  6.2 ±0.2 Na₂MoO₄ 0.12-0.36 0.18-0.31 .22-.28 0.25  0.25 ± .02 KH₂PO₄  85-255120-210 150-190 170   170 ± 2 FeSO₄ 27.0-84.0 40.0-70.0 50.0-60.0 55.7 55.7 ± 0.2 Na₂EDTA  37.0-112.0 55.0-94.0 67.0-82.0 74.6  74.6 ± 0.2Na₂H₂PO₄  85-255 120-210 150-190 170   170 ± 2 myo-Inositol  50-150 75-125  90-110 100   100 ± 2 Thiamine 0.2-0.6 0.3-0.5 0.36-0.44 0.4 0.4 ± 0.2 NAA 0.02-0.08 0.03-0.07 0.04-0.06 0.05  0.05 ± .02 BAP0.5-1.5 0.7-1.3 0.9-1.1 1    1 ± 0.2 Thidiazuron 0.2-0.8 0.3-0.7 0.4-0.60.5  0.5 ± 0.2 Meta-topolin 2.5-7.5 3.7-6.2 4.5-5.5 5    5 ± 2 Sugar g/L15-45 22-37 27-33 30   30 ± 2 Agar g/L 2.7-8.2 4.1-6.8 4.9-6.1 5.5  5.5± 0.2Media b-1(i-v):

Component b-1-i b-1-ii b-1-iii b-1-iv b-1-v NH₄NO₃  600-1800  900-15001080-1320 1200  1200 ± 2 Ca(NO₃)₂  838-2515 1257-2096 1510-1844 1677 1677 ± 2 K₂SO₄ 121-363 181-302 218-266 242   242 ± 2 MgSO₄ 270-830410-690 500-610 555   555 ± 2 MnSO₄ 12.60-38.00 19.00-31.70 22.80-27.8025.35 25.35 ± .02 ZnSO₄  6.4-19.5  9.6-16.2 11.5-14.0 12.9  12.9 ± 0.2CuSO₄ 0.018-0.055 0.027-0.046 0.033-0.041 0.037 0.037 ± 0.002 CaCl₂ 48-144  72-120  85-105 96   96 ± 2 H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2 6.2 ± 0.2 Na₂MoO₄ 0.12-0.36 0.18-0.31 .22-.28 0.25  0.25 ± .02 KH₂PO₄ 85-255 120-210 150-190 170   170 ± 2 FeSO₄ 27.0-84.0 40.0-70.050.0-60.0 55.7  55.7 ± 0.2 Na₂EDTA  37.0-112.0 55.0-94.0 67.0-82.0 74.6 74.6 ± 0.2 Na₂H₂PO₄  42-128  63-106 75-95 85   85 ± 2 myo-Inositol100-300 150-250 180-220 200   200 ± 2 Thiamine 0.4-1.4 0.6-1.1 0.8-1.00.9  0.9 ± 0.2 Pyridoxine 0.2-0.8 0.3-0.7 0.4-0.6 0.5  0.5 ± 0.2Nicotinic 0.2-0.8 0.3-0.7 0.4-0.6 0.5  0.5 ± 0.2 acid Glycine 1-31.5-2.5 1.75-2.25 2    2 ± 1 Riboflavin 10-30 15-25 18-22 20   20 ± 2BAP 0.1-0.3 0.15-0.25 0.17-0.22 0.2  0.2 ± 0.1 NAA 0.2-0.8 0.3-0.70.4-0.6 0.5  0.5 ± 0.2 Thidiazuron 0.36-1.12 0.56-0.94 0.67-.083 0.75 0.75 ± .02 2ip  7-23 11-19 13-17 15   15 ± 2 Sugar g/L 15-45 22-3727-33 30   30 ± 2 Carrageenan  3-11  4-10 5-8 7    7 ± 2 g/LMedia b-4(i-v):

Component b-4-i b-4-ii b-4-iii b-4-iv b-4-v NH₄NO₃  600-1800  900-15001080-1320 1200  1200 ± 2 Ca(NO₃)₂  838-2515 1257-2096 1510-1844 1677 1677 ± 2 K₂SO₄ 121-363 181-302 218-266 242   242 ± 2 MgSO₄ 270-830410-690 500-610 555   555 ± 2 MnSO₄ 12.60-38.00 19.00-31.70 22.80-27.8025.35 25.35 ± .02 ZnSO₄  6.4-19.5  9.6-16.2 11.5-14.0 12.9  12.9 ± 0.2CuSO₄ 0.018-0.055 0.027-0.046 0.033-0.041 0.037 0.037 ± .002 CaCl₂ 48-144  72-120  85-105 96   96 ± 2 H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2 6.2 ± 0.2 Na₂MoO₄ 0.12-0.36 0.18-0.31 .22-.28 0.25  0.25 ± .02 KH₂PO₄ 85-255 120-210 150-190 170   170 ± 2 FeSO₄ 27.0-84.0 40.0-70.050.0-60.0 55.7  55.7 ± 0.2 Na₂EDTA  37.0-112.0 55.0-94.0 67.0-82.0 74.6 74.6 ± 0.2 Na₂H₂PO₄  42-128  63-106 75-95 85   85 ± 2 myo-Inositol100-300 150-250 180-220 200   200 ± 2 Thiamine 0.4-1.4 0.6-1.1 0.8-1.00.9  0.9 ± 0.2 Pyridoxine 0.2-0.8 0.3-0.7 0.4-0.6 0.5  0.5 ± 0.2Nicotinic 0.2-0.8 0.3-0.7 0.4-0.6 0.5  0.5 ± 0.2 acid Glycine 1-31.5-2.5 1.75-2.25 2    2 ± 1 Riboflavin 10-30 15-25 18-22 20   20 ± 2BAP 2.5-7.5 3.7-6.2 4.5-5.5 5    5 ± 2 NAA 0.5-1.5 0.7-1.3 0.9-1.1 1   1 ± 0.2 Thidiazuron 0.36-1.12 0.56-0.94 0.67-.083 0.75  0.75 ± .022ip 10-30 15-25 18-22 20   20 ± 2 Sugar g/L 15-45 22-37 27-33 30   30 ±2 Carrageenan  3-11  4-10 5-8 7    7 ± 2 g/LMedia b-6(i-v):

Component b-6-i b-6-ii b-6-iii b-6-iv b-6-v NH₄NO₃  600-1800  900-15001080-1320 1200  1200 ± 2 Ca(NO₃)₂  838-2515 1257-2096 1510-1844 1677 1677 ± 2 K₂SO₄ 121-363 181-302 218-266 242   242 ± 2 MgSO₄ 270-830410-690 500-610 555   555 ± 2 MnSO₄ 12.60-38.00 19.00-31.70 22.80-27.8025.35 25.35 ± .02 ZnSO₄  6.4-19.5  9.6-16.2 11.5-14.0 12.9  12.9 ± 0.2CuSO₄ 0.018-0.055 0.027-0.046 0.033-0.041 0.037 0.037 ± .002 CaCl₂ 48-144  72-120  85-105 96   96 ± 2 H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2 6.2 ± 0.2 Na₂MoO₄ 0.12-0.36 0.18-0.31 .22-.28 0.25  0.25 ± .02 KH₂PO₄ 85-255 120-210 150-190 170   170 ± 2 FeSO₄ 27.0-84.0 40.0-70.050.0-60.0 55.7  55.7 ± 0.2 Na₂EDTA  37.0-112.0 55.0-94.0 67.0-82.0 74.6 74.6 ± 0.2 Na₂H₂PO₄  42-128  63-106 75-95 85   85 ± 2 myo-Inositol100-300 150-250 180-220 200   200 ± 2 Thiamine 0.4-1.4 0.6-1.1 0.8-1.00.9  0.9 ± 0.2 Pyridoxine 0.2-0.8 0.3-0.7 0.4-0.6 0.5  0.5 ± 0.2Nicotinic 0.2-0.8 0.3-0.7 0.4-0.6 0.5  0.5 ± 0.2 acid Glycine 1-31.5-2.5 1.75-2.25 2    2 ± 1 Riboflavin 10-30 15-25 18-22 20   20 ± 2NAA 0.5-1.5 0.7-1.3 0.9-1.1 1    1 ± 0.2 Thidiazuron 0.12-0.36 0.18-0.31.22-.28 0.25  0.25 ± .02 2ip 2.5-7.5 3.7-6.2 4.5-5.5 5    5 ± 2 Sugarg/L 12-37 15-35 20-30 25   25 ± 2 Agar g/L 2.7-8.2 4.1-6.8 4.9-6.1 5.5 5.5 ± 0.2 Carrageenan 1-3 1.5-2.5 1.75-2.25 2    2 ± 1 g/L

Note that for each of these media, its solid form is provided. Eachmedia can be transformed into a liquid media by removing agar orcarageenan and liquid forms of these media and their uses are expresslyincluded within the scope of the present disclosure.

The expected number of shoots may be different at different stages ofthe tissue culturing process and can also depend on the species ofbamboo. In general, however, at the beginning of the process,multiplication is from 1.0-2.0, 1.0-3.0 or 2.0-3.0 times. Onceestablished, multiplication can depend on the chosen container. Forexample, multiplication can range from, without limitation, 1-10 or 2-6plants per tube, 1-15 or 4-9 plants per jar, 1-20 or 9-17 plants per boxor 1-50 or 20-35 plants per jug. The number 1 is included becausecertain species or particular cell cultures require more time in Stage 1or Stage 2 media before multiplication begins. By carrying them throughthe process, however, most if not all begin multiplication within anumber of cycles. For example, some cell cultures may begin to multiplyonly after 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or18 months in culture.

Methods disclosed herein can produce the following non-limiting numberof shoots from a single explant: 100, 500, 1,000; 5,000; 10,000, 20,000,50,000, 100,000, 250,000, 500,000, 750,000, 1,000,000 or more.

Following multiplication through culturing and subculturing, particularplant shoots can be selected for transition to ex vitro conditions.Generally, media that support transition to ex vitro conditionsrepresent a Stage 2, Stage 3, Stage 4 or Stage 5 media. Non-limitingexamples of such media include:

Media Ech(i-v):

Ech- Component Ech-i Ech-ii Ech-iii iv Ech-v NH₄NO₃  825-2475 1237-20631485-1815 1650  1650 ± 2 KNO₃  950-2850 1425-2375 1710-2090 1900  1900 ±2 MgSO₄ 185-555 275-465 330-410 370   370 ± 2 MnSO₄  8.0-26.0 12.0-22.015.0-19.0 16.9  16.9 ± 0.2 ZnSO₄  4.0-12.0  6.0-10.0 8.0-9.0 8.6  8.6 ±0.2 CuSO₄ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ± .002 CaCl₂220-660 330-350 400-480 440   440 ± 2 KI 0.40-1.25 0.60-1.05 0.75-0.900.83  0.83 ± .02 CoCl₂ 0.012-0.378 0.020-0.030 0.022-0.028 0.025 0.025 ±.002 H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2  6.2 ± 0.2 Na₂MoO₄ 0.12-0.360.18-0.31 .22-.28 0.25  0.25 ± .02 KH₂PO₄  85-255 120-210 150-190 170  170 ± 2 FeSO₄ 13.0-42.0 20.8-34.7 25.1-30.5 27.8  27.8 ± 0.2 Na₂EDTA18.6-56.0 28.0-46.6 33.6-41.0 37.3  37.3 ± 0.2 myo-Inositol  50-150 75-125  90-110 100   100 ± 2 Thiamine 0.2-0.6 0.3-0.5 0.35-0.45 0.4 0.4 ± 0.2 Pyridoxine 0.2-0.8 0.3-0.7 0.4-0.6 0.5  0.5 ± 0.2 Nicotinic0.2-0.8 0.3-0.7 0.4-0.6 0.5  0.5 ± 0.2 acid Glycine 1-3 1.5-2.51.75-2.25 2    2 ± 1 NAA 0.05-0.15 0.07-0.12 0.09-0.11 0.1  0.1 ± 0.02IAA 0.02-0.08 0.03-0.07 0.04-0.06 0.05  0.05 ± .02 Sugar g/L 15-45 22-3727-33 30   30 ± 2 Agar g/L 2.7-8.2 4.1-6.8 4.9-6.1 5.5  5.5 ± 0.2Media BR-2(i-v):

BR-2- Component BR-2-i BR-2-ii BR-2-iii iv BR-2-v NH₄NO₃  700-21001050-1750 1260-1540 1400   1400 ± 2 Ca(NO₃)₂  973-2919 1459-24331752-2140 1946   1946 ± 2 K₂SO₄  606.3-1818.8  909.4-1515.61091.5-1333.5 1212.5 1212.5 ± 0.2 MgSO₄  370-1110 555-925 665-815 740  740 ± 2 MnSO₄ 16.9-50.7 25.4-42.3 30.5-37.1 33.8  33.8 ± 0.2 ZnSO₄ 8.6-25.8 12.9-21.5 15.5-18.0 17.2  17.2 ± 0.2 CuSO₄ 0.02-0.08 0.03-0.070.04-0.06 0.05  0.05 ± .02 CaCl₂  72-216 108-180 130-158 144   144 ± 2H₃BO₃ 3.0-9.0 4.0-8.0 5.0-7.0 6.2   6.2 ± 0.2 Na₂MoO₄ 0.12-0.360.18-0.31 .22-.28 0.25  0.25 ± .02 KH₂PO₄  72-342 202-338 243-297 270  270 ± 2 FeSO₄ 16.68-50.04 25.02-41.70 30.06-36.66 33.36  33.36 ± .02Na₂EDTA 22.38-67.14 33.57-55.95 40.36-49.16 44.76  44.76 ± .02myo-Inositol 100-300 150-250 180-220 200   200 ± 2 Thiamine 0.4-1.40.6-1.2 0.8-1.0 0.9   0.9 ± 0.2 Pyridoxine 0.2-0.8 0.3-0.7 0.4-0.6 0.5  0.5 ± 0.2 Nicotinic 0.2-0.8 0.3-0.7 0.4-0.6 0.5   0.5 ± 0.2 acidGlycine 1-3 1.5-2.5 1.75-2.25 2    2 ± 1 Riboflavin 10-30 15-25 18-22 20   20 ± 2 Ascorbic  50-150  75-125  90-110 100   100 ± 2 Acid NAA0.2-0.8 0.3-0.7 0.4-0.6 0.5   0.5 ± 0.2 Sugar g/L 15-45 22-37 27-33 30   30 ± 2 Carrageenan  4-12  6-10 7-9 8    8 ± 2 g/L Charcoal g/L150-450 220-370 270-330 300   300 ± 2

Media Amel(i-v):

Amel- Component Amel-i Amel-ii Amel-iii iv Amel-v NH₄NO₃  410-1240 620-1030 740-910 825   825 ± 2 Ca(NO₃)₂  475-1425  710-1190  855-1045950   950 ± 2 MgSO₄  90-280 140-230 160-200 185   185 ± 2 MnSO₄ 4.20-12.70  6.30-10.60 7.65-9.25 8.45  8.45 ± .02 ZnSO₄ 2.0-6.5 3.0-5.53.5-5.0 4.3   4.3 ± 0.2 CuSO₄ .0062-.0188 .0094-.0156 .0115-.0135 0.01250.0125 ± .0002 CaCl₂ 110-330 165-285 195-240 220   220 ± 2 KI .207-.623.310-.520 .375-.455 0.415  0.415 ± .002 H₃BO₃ 1.5-4.6 2.3-4.0 2.8-3.43.1   3.1 ± 0.2 CaCl₂ .0062-.0188 .0094-.0156 .0115-.0135 0.0125 0.0125± .0002 Na₂MoO₄ .062-.188 .093-.157 .115-.135 0.125  0.125 ± .002 KH₂PO₄ 40-130  60-110 75-95 85    85 ± 2 FeSO₄  6.8-20.9 10.4-17.5 12.5-15.513.9  13.9 ± 0.2 Na₂EDTA  9.32-27.98 13.95-23.35 16.85-20.45 18.65 18.65 ± .02 Na₂H₂PO₄  40-130  60-110 75-95 85    85 ± 2 myo-Inositol 50-150  75-125  90-110 100   100 ± 2 Thiamine 0.2-0.6 0.3-0.5 0.36-0.440.4   0.4 ± .2 Pyridoxine 0.2-0.8 0.3-0.7 0.4-0.6 0.5   0.5 ± 0.2Nicotinic 1-3 1.5-2.5 1.75-2.25 2    2 ± 1 acid Riboflavin 2.5-7.53.7-6.2 4.5-5.5 5    5 ± 2 NAA 0.2-0.8 0.3-0.7 0.4-0.6 0.5   0.5 ± 0.2IBA 0.5-1.5 0.7-1.3 0.9-1.1 1    1 ± 0.5 Sugar g/L 15-45 22-37 27-33 30   30 ± 2 Agar g/L 1.5-4.5 2.0-4.0 2.5-3.5 3    3 ± 2 Carrageenan2.5-7.5 3.7-6.2 4.5-5.5 5    5 ± 2 g/L Charcoal g/L 2.5-7.5 3.7-6.24.5-5.5 5    5 ± 2

During transition to ex vitro conditions, shoots and media can be placedin air permeable or air impermeable containers.

Each of the media described herein can be used in combination with eachother media in a method, system or kit described herein. Moreover, themedia can be combined in combinations greater than two (e.g., a kit mayinclude 2 of the different media provided herein, or include 3 of thedifferent media provided herein, or include more than 3 of the differentmedia described herein). While not explicitly describing each possiblecombination herein, one of ordinary skill in the art should understandthat this disclosure supports all possible combinations.

Following transition to ex vitro conditions, but before plants areplaced in soil, or exposed to less regulated growing conditions, theplants can undergo a series of treatments designed to acclimate them toan unregulated growing environment. This is because some plants, whenmicrocultured, do not develop adequate defensive structures, such aswaxy cuticles to protect them from ordinary environmental conditions.The treatments that plants can undergo prior to being placed in anunregulated environment can include, without limitation, acclimatizationto humidity, acclimatization to variations in temperature, andacclimatization to wind pressure. These acclimatization factors can beintroduced gradually and/or in a staggered fashion.

Representative genus of bamboo appropriate for use with the disclosuresherein include: Acidosasa; Ampelocalamus; Arundinaria; Bambusa;Bashania; Borinda; Brachystachyum; Cephalostachyum; Chimonobambusa;Chimonocalamus; Chusquea; Dendrocalamus; Dinochloa; Drepanostachyum;Eremitis; Fargesia; Gaoligongshania; Gigantochloa; Guadua;Hibanobambusa; Himalayacalamus; Indocalamus; Indosasa; Lithachne;Melocalamus; Melocanna; Menstruocalamus; Nastus; Neohouzeaua; Neololeba;Ochlandra; Oligostachyum; Olmeca; Otatea; Oxytenanthera; Phyllostachys;Pleioblastus; Pseudosasa; Raddia; Rhipidocladum; Sarocalamus; Sasa;Sasaella; Sasamorpha; Schizostachyum; Semiarundinaria; Shibataea;Sinobambusa; Thamnocalamus; Thyrsostachys; and Yushania.

Non-limiting examples of species within these genus include:

Acidosasa: Edulis

Ampelocalamus: Scandens

Arundinaria: Arundinaria appalachiana; Arundinaria funghomii;Arundinaria gigantea; Arundinaria gigantea ‘Macon’; and ArundinariaTecta

Bambusa: arnhemica; balcooa; bambos; basihirsuta; beecheyana; beecheyanavar pubescens; blumeana; boniopsis; burmanica; chungii; chungii var.Barbelatta; cornigera; dissimulator; dissimulator ‘Albinodia’; distegia;dolichoclada; dolichoclada ‘Stripe’; dolichomerithalla ‘Green stripe’;dolichomerithalla ‘Silverstripe’; emeiensis ‘Chrysotrichus’; emeiensis‘Flavidovirens’; emeiensis ‘Viridiflavus’; eutuldoides; eutuldoides‘Viridivittata’; gibba; glaucophylla; intermedia; lako; lapidea;longispiculata; maculata; malingensis; multiplex; multiplex ‘AlphonseKarr’; multiplex ‘Fernleaf Stripestem’; multiplex ‘Fernleaf’; multiplex‘Golden Goddess’; multiplex ‘Goldstripe’; multiplex ‘Midori Green’;multiplex ‘Riviereorum’; multiplex ‘Silverstripe’; multiplex ‘Tiny FernStriped’; multiplex ‘Tiny Fern’; multiplex ‘Willowy’; nutans; odashimae;odashimae X B. Tuldoides; oldhamii; oliveriana; pachinensis;pervariabilis; pervariabilis ‘Viridistriatus’; rigida; rutila;sinospinosa; sp ‘Hirose’; sp. ‘Clone X’; sp. ‘Nana’; sp. ‘Polymorpha’;sp. ‘Richard Waldron’; stenostachya; suberecta; textilis; textilis‘Dwarf’; textilis ‘Kanapaha’; textilis ‘Maculata’; textilis ‘Mutabilis’;textilis ‘Scranton’; textilis var. Albostriata; textilis var. Glabra;textilis var. Gracilis; tulda; tulda ‘Striata’; Tuldoides; variostriata;ventricosa; ventricosa ‘Kimmei’; vulgaris; vulgaris ‘Vittata’; vulgaris‘Wamin Striata’; and vulgaris ‘Wamin’

Bashania: Fargesii; and Qingchengshanensis

Borinda: KR 5288; Albocerea; Angustissima; Contracta; Frigidorum;Fungosa; fungosa ‘White Cloud’; Lushuiensis; Macclureana; Nujiangensis;Papyrifera; Perlonga; sp. ‘Muliensis’; and Yulongshanensis

Brachystachyum: densiflorum; and densiflorum var. villosum

Cephalostachyum: Pergracile; and Virgatum

Chimonobambusa: macrophylla ‘Intermedia’; Marmorea; marmorea‘Variegata’; Quadrangularis; quadrangularis ‘Joseph de Jussieu’;quadrangularis ‘Suow’; quadrangularis ‘Yellow Groove’; Szechuanensis;and Tumidissinoda

Chimonocalamus: Pallens

Chusquea: Andina; Circinata; circinata ‘Chiapas’ Coronalis; Culeou;culeou ‘Argentina’; culeou ‘Caña Prieta’; culeou ‘Hillier's Form’;Cumingii; Delicatula; Foliosa; Galeottiana; Gigantea; Glauca;Liebmannii; Macrostachya; mimosa ssp. Australis; Montana; Muelleri;Pittieri; Simpliciflora; sp. ‘Chiconquiaco’; sp. ‘Las Vigas’; Subtilis;Sulcata; Tomentosa; Uliginosa; Valdiviensis; and Virgata

Dendrocalamus: Asper; asper ‘Betung Hitam’; Brandisii; brandisii‘Black’; brandisii (variegated); Calostachyus; Giganteus; giganteus(Quail Clone); giganteus (variegated); Hamiltonii; Jianshuiensis;jianshuiensis (variegated); Latiflorus; latiflorus ‘Mei-nung’;Membranaceus; Minor; minor ‘Amoenus’; Sikkimensis; Sinicus; sp.‘Maroochy’; sp. ‘Parker's Giant’; Strictus; Validus; and Yunnanicus

Dinochloa: Malayana; and Scandens

Drepanostachyum: falcatum var. sengteeanum; and Khasianum

Eremitis: Eremitis

Fargesia: Adpressa; Apircirubens; apircirubens ‘White Dragon’; Denudata;dracocephala ‘Rufa’; Murieliae; murieliae ‘SABE 939’; murieliae‘Vampire’; murieliae (next generation seedlings); Nitida; nitida‘Jiuzhaigou’; Robusta; robusta ‘Campbell’; robusta ‘Wolong’; sp.‘Scabrida’; and Utilis

Gaoligongshania: Gaoligongshania and Megalothyrsa

Gigantochloa: Hitam Hijau; Albociliata; Apus; Atroviolacea; Atter;Hasskarliana; Levis; Luteostriata; Maxima; Pseudoarundinacea; Ridleyi;Robusta; sp ‘Rachel Carson’; sp. ‘Bali White Stripe’; sp. ‘Sumatra3751’; sp. ‘Widjaja 3827’; and Wrayii

Guadua: Amplexifolia; Angustifolia; angustifolia ‘Bicolor’; angustifolia‘Less Thorny’; Chacoensis; Longifolia; Paniculata; sp. ‘Aureocaulis’;and Velutina

Hibanobambusa: Tranquillans; and tranquillans ‘Shiroshima’

Himalayacalamus: Falconeri; falconeri ‘Damarapa’; Hookerianus; Planatus;and Porcatus

Indocalamus: Cordatus; Latifolius; latifolius ‘Hopei’; Longiauritus; sp.‘Hamadae’; sp. ‘Solidus’; Tessellatus; and Victorialis

Indosasa: Crassiflora; and Gigantea

Lithachne: Humilis

Melocalamus: Arrectus

Melocanna: Baccifera

Menstruocalamus: Sichuanensis

Nastus: Elatus

Neohouzeaua: Mekongensis

Neololeba: Atra

Ochlandra: Stridula

Oligostachyum: Glabrescens

Olmeca: Recta

Otatea: acuminata ‘Michoacan’; acuminata ssp. Acuminata; acuminata ssp.Aztecorum, acuminata ssp. aztecorum ‘Dwarf’; Fimbriata; and glauca‘Mayan Silver’

Oxytenanthera: Abyssinica; and Braunii

Phyllostachys: Acuta; Angusta; Arcana; arcana ‘Luteosulcata’;Atrovaginata; Aurea; aurea ‘Albovariegata’; aurea ‘Dr Don’; aurea‘Flavescens-inversa’; aurea ‘Holochrysa’; aurea ‘Koi’; aurea‘Takemurai’; Aureosulcata; aureosulcata ‘Alata’; aureosulcata‘Aureocaulis’; aureosulcata ‘Harbin Inversa’; aureosulcata ‘Harbin’;aureosulcata ‘Pekinensis’; aureosulcata ‘Spectabilis’; Aurita;Bambusoides; bambusoides ‘Albovariegata’; bambusoides ‘CastillonInversa’; bambusoides ‘Castillon’; bambusoides ‘Golden Dwarf’;bambusoides ‘Job's Spot’; bambusoides ‘Kawadana’; bambusoides ‘Marliac’;bambusoides ‘Rib Leaf’; bambusoides ‘Richard Haubrich’; bambusoides‘Slender Crookstem’; bambusoides ‘Subvariegata’; bambusoides ‘Tanakae’;bambusoides ‘White Crookstem’; Bissetii; bissetii ‘Dwarf’; Dulcis;Edulis; edulis ‘Anderson’; edulis ‘Bicolor’; edulis ‘Goldstripe’; edulis‘Heterocycla’; Elegans; Flexuosa; flexuosa ‘Kimmei’; Glauca; glauca‘Notso’; glauca ‘Yunzhu’; Heteroclada; heteroclada ‘Purpurata’;heteroclada ‘Solidstem’; Hispida; Humilis; Incamata; Iridescens;Kwangsiensis; Lithophila; Lofushanensis; Makinoi; mannii ‘Decora’;mannii ‘Mannii’; Meyeri; Nidularia; nidularia ‘Farcta’; nidularia‘Smoothsheath’; Nigra; nigra ‘Bory’; nigra ‘Daikokuchiku’; nigra ‘Hale’;nigra ‘Henon’; nigra ‘Megurochiku’; nigra ‘Mejiro’; nigra ‘Muchisasa’;nigra ‘Othello’; nigra ‘Punctata’; nigra ‘Shimadake’; nigra ‘Tosaensis’;Nuda; nuda ‘Localis’; Parvifolia; Platyglossa; Praecox; praecox‘Prevernalis’; praecox ‘Viridisulcata’; Prominens; Propinqua; propinqua‘Beijing’; Robustiramea; Rubromarginata; Stimulosa; Varioauriculata;Violascens; Viridiglaucescens; Viridis; viridis ‘Houzeau’; viridis‘Robert Young’; Vivax; vivax ‘Aureocaulis’; vivax ‘Black Spot’; vivax‘Huangwenzhu Inversa’; and vivax ‘Huangwenzhu’

Pleioblastus: Akebono; Amarus; Argenteostriatus; Chino; chino‘Angustifolia’; chino ‘Elegantissimus’; chino ‘Kimmei’; chino‘Murakamiansus’; chino ‘Vaginatus Variegatus’; Distichus; distichus‘Mini’; Fortunei; Gauntlettii; Gramineus; gramineus ‘Monstrispiralis’;Hindsii; Humilis; humilis ‘Albovariegatus’; humilis ‘Variegatus’;Juxianensis; Kodzumae; Kongosanensis; kongosanensis ‘Akibensis’;kongosanensis ‘Aureostriatus’; Linearis; linearis ‘Nana’; Nagashima;Oleosus; Pygmaeus; pygmaeus ‘Greenstripe’; pygmaeus ‘Ramosissimus’;shibuyanus ‘Tsuboi’; Simonii; simonii ‘Variegatus’; Viridistriatus;viridistriatus ‘Chrysophyllus’; and Xestrophyllus

Pseudosasa: Amabilis; Cantori; Guanxianensis; Japonica; japonica‘Akebono’; japonica ‘Akebono-suji’; japonica ‘Pleioblastoides’; japonica‘Tsutsumiana’; japonica ‘Variegata’; Longiligula; Owatarii; Usawai; andViridula

Raddia: Brasiliensis; and Distichophylla

Rhipidocladum: Pittieri; and Racemiflorum

Sarocalamus: Faberi; and Fangianus

Sasa: Cernua; Gracillima; Hayatae; Kagamiana; kagamiana ssp. Yoshinoi;Kurilensis; kurilensis ‘Shimofuri’; Megalophylla; Nagimontana; nipponica(hort.); Oshidensis; Palmata; Senanensis; Shimidzuana; sp. Tsuboiana;and Veitchii

Sasaella: Bitchuensis; hidaensis ‘muraii’; Masamuneana; masamuneana‘Albostriata’; masamuneana ‘Aureostriata’; Ramosa; Sasakiana; andShiobarensis

Sasamorpha: Borealis

Schizostachyum: Brachycladum; brachycladum ‘Bali Kuning’; Caudatum;Glaucifolium; Jaculans; Lima; and sp. ‘Murray Island’

Semiarundinaria: Fastuosa; fastuosa ‘Viridis’; Fortis; Kagamiana;Lubrica; Makinoi; Okuboi; sp. Maruyamana; sp. ‘Korea’; Yashadake;yashadake ‘Kimmei’; and yashadake ‘kimmei inversa’

Shibataea: Chinensis; Kumasaca; kumasaca ‘Albostriata’; kumasaca‘Aureostriata’; Lancifolia; and Nanpingensis

Sinobambusa: Gigantea; Intermedia; Tootsik; and tootsik ‘Albostriata’

Thamnocalamus: aristatus ‘Aristatus hort. US’; Crassinodus; crassinodus‘Kew Beauty’; crassinodus ‘Mendocino’; crassinodus ‘Merlyn’; nepalensis‘Nyalam’; Spathiflorus; and Tessellatus

Thyrsostachys: Oliveri; and Siamensis

Yushania: Alpina; Anceps; anceps ‘Pitt White’; Boliana; Brevipaniculata;Exilis; Maculata; and maling

Particularly useful species include: edulis; scandens; ArundinariaGigantea; Arundinaria Tecta; Bambusa Balcooa; Bambusa Bambos; BambusaOldhamii; Bambusa Textilis; Bambusa Tulda; Bashania Fargesii;Brachystachyum Densiflorum; Chusquea Gigantea; Dendrocalamus Asper;Dendrocalamus Brandisii; Dendrocalamus Giganteus; DendrocalamusHamiltonii; Dendrocalamus Strictus; Fargesia Denudata; Fargesiadracocephala ‘Rufa’; Fargesia Murieliae; Fargesia Nitida; FargesiaRobusta; Fargesia robusta ‘Wolong’; Fargesia sp. ‘Scabrida’; GuaduaAmplexifolia; Guadua Paniculata; Himalayacalamus Falconeri; IndocalamusTessellatus; Ochlandra Stridula; Otatea acuminata ssp. Aztecorum;Phyllostachys Atrovaginata; Phyllostachys Aurea; PhyllostachysBambusoides; Phyllostachys Bissetii; Phyllostachys Edulis; Phyllostachysedulis ‘Heterocycla’; Phyllostachys Glauca; Phyllostachys Iridescens;Phyllostachys Kwangsiensis; Phyllostachys Nidularia; PhyllostachysNigra; Phyllostachys nigra ‘Henon’; Phyllostachys Nuda; PhyllostachysParvifolia; Phyllostachys Praecox; Phyllostachys Propinqua;Phyllostachys Viridis; Phyllostachys Vivax; Pleioblastus Distichus;Pleioblastus Fortunei; Pleioblastus Linearis; Pseudosasa Japonica; SasaKurilensis; Sasa Veitchii; Sasaella Masamuneana; Sasamorpha Borealis;Schizostachyum Brachycladum; Schizostachyum brachycladum ‘Bali Kuning’;Schizostachyum Caudatum; Schizostachyum Glaucifolium; SchizostachyumJaculans; Schizostachyum Lima; Schizostachyum sp. ‘Murray Island’;Semiarundinaria Fastuosa; Semiarundinaria Yashadake; Shibataea Kumasaca;Sinobambusa Gigantea; Thamnocalamus Crassinodus; ThamnocalamusTessellatus; Yushania Alpina; and Yushania maling.

As one of ordinary skill in the art appreciates, many species of bamboohave different common names. Accordingly, the following terminology andlanguage comparisons are provided.

Classic Name Equivalent ACIDOSASA gigantea INDOSASA giganteaARTHROSTYLIDIUN sp. (hort) CHUSQUEA circinata ARUNDINARIA alpinaYUSHANIA alpina ARUNDINARIA amabilis PSEUDOSASA amabilis ARUNDINARIAanceps YUSHANIA anceps ARUNDINARIA auricoma PLEIOBLASTUS viridistriatusARUNDINARIA falconeri DREPANOSTACHYUM falcatum ‘var. sengteeanum’ARUNDINARIA fangiana SAROCALAMUS fangianus ARUNDINARIA hookerianusHIMALAYACALAMUS hookerianus ARUNDINARIA macrosperma ARUNDINARIA giganteaARUNDINARIA maling YUSHANIA maling ARUNDINARIA tessellata THAMNOCALAMUStessellatus ARUNDINARIA vagans SASAELLA ramosa BAMBUSA arundinaceaBAMBUSA bambos BAMBUSA dissemulator BAMBUSA dissimulator BAMBUSA edulisBAMBUSA odashimae BAMBUSA forbesii NEOLOLEBA atra BAMBUSA glaucescensBAMBUSA multiplex BAMBUSA multiplex ‘Green Alphonse’ BAMBUSA multiplex‘Midori Green’ Bambusa mutabilis Bambusa textilis ‘Mutabilis’ Bambusatuldoides ‘Clone X’ Bambusa sp. ‘Clone X’ Bambusa tuldoides ‘ventricosa’Bambusa ventricosa Bambusa variegata (hort.) Bambusa glaucophyllaBAMBUSA vulgaris ‘Striata’ BAMBUSA vulgaris ‘Vittata’ BASHANIA faberiSAROCALAMUS faberi Borinda boliana Yushania boliana CHIMONOBAMBUSAfalcata HIMALYACALAMUS hookerianus CHUSQUEA breviglumis CHUSQUEAgigantea Chusquea breviglumis Chusquea culeou CHUSQUEA nigricansCHUSQUEA culeou ‘Ca?a Prieta’ CHUSQUEA quila CHUSQUEA valdiviensisDENDROCALAMUS affinis BAMBUSA emeiensis DENDROCALAMUS membranaceusBAMBUSA membranacea DREPANOSTACHYUM falcatum HIMALAYACALAMUS hookerianusDREPANOSTACHYUM falconeri DREPANOSTACHYUM falcatum ‘var. sengteeanum’DREPANOSTACHYUM hookerianum HIMALAYACALAMUS falconeri ‘Damarapa’DREPANOSTACHYUM sengteeanum HIMALAYACALAMUS falconeri FARGESIAangustissima BORINDA angustissima FARGESIA crassinodus THAMNOCALAMUScrassinodus Fargesia dracocephala Fargesia apircirubens Fargesiadracocephala ‘White Dragon’ Fargesia apircirubens ‘White Dragon’FARGESIA frigida BORINDA frigidorum FARGESIA fungosa BORINDA fungosaFARGESIA sp “A-4” FARGESIA adpressa Fargesia sp. ‘rufa’ Fargesiadracocephala ‘Rufa’ GELIDOCALAMUS fangianus SAROCALAMUS fangianusGIGANTOCHLOA atroviolacea ‘Timor BAMBUSA lako Black’ GIGANTOCHLOAluteostriata BAMBUSA luteostriata GIGANTOCHLOA verticillata GIGANTOCHLOApseudoarundinacea Himalayacalamus asper Himalayacalamus planatusHIMALAYACALAMUS falconeri DREPANOSTACHYUM falcatum ‘var. ‘glomeratum’sengteeanum’ HIMALAYACALAMUS intermedius Yushania bolianaHIMALAYACALAMUS planatus HIMALAYACALAMUS asper (hort.) HIMALAYACALAMUSplanatus Neomicrocalamus microphyllus (hort.) NEOMICROCALAMUSmicrophyllus HIMALAYACALAMUS planatus NEOSINOCALAMUS affinis BAMBUSAemeiensis ‘Chrysotrichus’ Otatea acuminata ‘Mayan Silver’ Otatea glauca‘Mayan Silver’ OTATEA aztecorum OTATEA acuminata ssp. aztecorumPHYLLOSTACHYS cerata PHYLLOSTACHYS heteroclada PHYLLOSTACHYS congestaPHYLLOSTACHYS atrovaginata PHYLLOSTACHYS decora PHYLLOSTACHYS mannii‘Decora’ PHYLLOSTACHYS heterocycla PHYLLOSTACHYS edulis ‘Heterocycla’PHYLLOSTACHYS heterocycla PHYLLOSTACHYS edulis pubescens PHYLLOSTACHYSheterocycla PHYLLOSTACHYS edulis ‘Anderson’ pubescens ‘Anderson’PHYLLOSTACHYS purpurata PHYLLOSTACHYS heteroclada ‘Purpurata’PHYLLOSTACHYS purpurata PHYLLOSTACHYS heteroclada ‘Solidstem’‘Solidstem’ PHYLLOSTACHYS purpurata PHYLLOSTACHYS heteroclada‘Straightstem’ PLEIOBLASTUS akibensis PLEIOBLASTUS kongosanensis‘Akibensis’ PLEIOBLASTUS gramineus ‘Raseetsuchiku’ PLEIOBLASTUSgramineus ‘Monstrispiralis’ PLEIOBLASTUS variegatus PLEIOBLASTUSfortunei Qiongzhuea tumidissinoda Chimonobambusa tumidissinoda SASAasahinae SASA shimidzuana SASA humilis PLEIOBLASTUS humilis SASA pygmaeaPLEIOBLASTUS pygmaeus SASA tessellata INDOCALAMUS tessellatus SASAvariegata PLEIOBLASTUS fortunei Sasa veitchii ‘Minor’ Sasa hayataeSASAELLA glabra ‘Albostriata’ SASAELLA masamuneana ‘Albostriata’SASAELLA masamuneana rhyncantha SASAELLA masamuneana SASAELLA rhyncanthaSASAELLA masamuneana SEMIARUNDINARIA villosa SEMIARUNDINARIA okuboiSINARUNDINARIA FARGESIA TETRAGONOCALAMUS angulatus CHIMONOBAMBUSAquadranqularis THAMNOCALAMUS spathaceus FARGESIA murieliae YUSHANIAaztecorum OTATEA acuminata ssp. aztecorum

Chinese & Japanese Names

Chinese Botanical Japanese Botanical Cha Gang zhu Pseudosasa HachikuPhyllostachys nigra amabilis ‘Henon’ Che Tong zhu Bambusa HoteichikuPhyllostachys aurea sinospinosa Fang zhu Chimonobambusa KikkochikuPhyllostachys edulis quadrangularis ‘Heterocycla’ Fo du zhu BambusaKumazasa Sasa veitchii (not ventricosa Shibataea kumasaca) Gui zhuPhyllostachys Kurochiku Phyllostachys nigra bambusoides Han zhuChimonobambusa Madake Phyllostachys marmorea bambusoides Hong Bian zhuPhyllostachys Medake Pleioblastus simonii rubromarginata Hou zhuPhyllostachys Moso Phyllostachys edulis nidularia Hui Xiang zhuChimonocalamus Narihira Semiarundinaria pallens fastuosa Jin zhuPhyllostachys Okame-zasa Shibataea kumasaca sulphurea Ma zhuDendrocalamus Yadake Pseudosasa japonica latiflorus Mao zhuPhyllostachys edulis Qiong zhu Chimonobambusa tumidissinoda Ren Mian zhuPhyllostachys aurea Shui zhu Phyllostachys heteroclada Wu Ya zhuPhyllostachys atrovaginata Xiang Nuo zhu Cephalostachyum pergracile Zizhu Phyllostachys nigra

English Names

English Botanical Arrow Pseudosasa japonica Beechey Bambusa beecheyanaBlue Himalayacalamus hookerianus Black Phyllostachys nigra Buddha'sBambusa Belly ventricosa Candy Stripe Himalayacalamus or Candy falconericane ‘Damarapa’ Canebrake Arundinaria gigantea Chinese Bambusa multiplexGoddess ‘Riviereorum’ Chinese Bambusa Thorny sinospinosa Common Bambusavulgaris Dwarf Fern Pleioblastus Leaf distichus Dwarf PleioblastusWhitestripe fortunei Fernleaf Bambusa multiplex ‘Fernleaf’ FountainFargesia nitida Giant Thorny Bambusa bambos Giant Timber Bambusaoldhamii Green Yushania alpina Mountain Golden Phyllostachys aureaGolden Phyllostachys Golden aurea ‘Holochrysa’ “Heavenly not a bambooBamboo” (Nandina domestica) Hedge Bambusa multiplex HimalayanHimalayacalamus Blue hookerianus Horsehoof Bambusa lapidea Iron RangeNeololeba atra Japanese Phyllostachys Timber bambusoides “Lucky Dracaenasanderiana ¹ Bamboo” Male Dendrocalamus strictus Marbled Chimonobambusamarmorea Mexican Otatea acuminata Weeping subsp. aztecorum MonasteryThyrsostachys siamensis Oldham's Bambusa oldhamii Painted Bambusavulgaris ‘Vittata’ Punting Pole Bambusa tuldoides River Cane Arundinariagigantea Square Chimonobambusa quadrangularis Stone Phyllostachysangusta & P. nuda Sweetshoot Phyllostachys dulcis Switch CaneArundinaria tecta Tea Stick Pseudosasa amabilis Temple Semiarundinariafastuosa Timor Black Bambusa lako Tonkin Cane Pseudosasa amabilisTortoise Phyllostachys edulis Shell ‘Heterocycla’ Tropical GigantochloaBlack atroviolacea Umbrella Fargesia murieliae Water Phyllostachysheteroclada Weaver's Bambusa textilis Wine Oxytenanthera braunii YellowPhyllostachys Groove aureosulcata ¹Technically not a bamboo but includedwithin the meaning of bamboo herein.

By means of the media, systems and methods described and disclosedherein, it is possible for one of ordinary skill in the art to achieverolling tissue cultures of bamboo. As used herein, “rolling tissueculture” means that the multiplication process can continuesubstantially indefinitely by continuing to separate and multiplyshoots. In one embodiment, one shoot is placed in a tube and the shootmultiplies into a number of additional shoots. After multiplication,each shoot or a subset of the shoots are separated and each placed in asubsequent tube for further multiplication. This process can continuewhile at various times, some or all shoots can be removed from themultiplication process and transitioned to ex vitro conditions. Bycontinuing indefinitely, it is meant that 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, etc. daymultiplication cycles can be repeated without initiating new explantsfor at least 1 month, for at least 3 months, for at least 6 months, forat least 9 months, for at least 12 months, for at least 15 months, forat least 18 months, for at least 21 months, for at least 24 months orfor at least 36 months. Particular ranges of days in multiplicationcycles include 10-120 days; 10-100 days; 10-80 days; 10-60 days; 10-42days; 10-40 days; 10-20 days; 14-120 days; 14-90 days; 14-70 days;14-50; 14-42 days; 14-30 days; 14-21 days; 12-42 days; 20-60 days; 10-15days; 14-20 days; 14-18 days etc.

These media systems and methods can be packaged and/or described invarious kits. Kits can include, without limitation, one or more of thefollowing in a package or container: (1) one or more media; and (2) oneor more explants from one or more species of bamboo. In certainnon-limiting embodiments, the media can be b-9-i media, b-9-ii media,b-9-iii media, b-9-iv media, b-9-v media, CW2-i media, CW2-ii media,CW2-iii media, CW2-iv media, CW2-v media, b-10-i media, b-10-ii media,b-10-iii media, b-10-iv media, b-10-v media, b-11-i media, b-11-iimedia, b-11-iii media, b-11-iv media, b-11-v media, b-12c-i media,b-12c-ii media, b-12c-iii media, b-12c-iv media, b-12c-v media, b-1-imedia, b-1-ii media, b-1-iii media, b-1-iv media, b-1-v media, b-4-imedia, b-4-ii media, b-4-iii media, b-4-iv media, b-4-v media, b-6-imedia, b-6-ii media, b-6-iii media, b-6-iv media, b-6-v media, CW1-imedia, CW1-ii media, CW1-iii media, CW1-iv media, CW1-v media, CW3-imedia, CW3-ii media, CW3-iii media, CW3-iv media, CW3-v media, CW4-imedia, CW4-ii media, CW4-iii media, CW4-iv media, CW4-v media, CW5-imedia, CW5-ii media, CW5-iii media, CW5-iv media, CW5-v media, CW6-imedia, CW6-ii media, CW6-iii media, CW6-iv media, CW6-v media, Br-2-imedia, Br-2-ii media, Br-2-iii media, Br-2-iv media, Ech-i media, Ech-iimedia, Ech-iii media, Ech-iv, Amel-i media, Amel-ii media, Amel-iiimedia, Amel-iv media or Amel-v. In another embodiment, the kits cancomprise one or more containers for the tissue culturing processincluding without limitation, tubes, jars, boxes or jugs. In anotherembodiment the kits can comprise instructions for the tissue culturingof bamboo. In another embodiment, the kits comprise combinations of theforegoing. Components of various kits can be found in the same ordifferent containers. Additionally, when a kit is supplied, thedifferent components of the media can be packaged in separate containersand admixed immediately before use. Such packaging of the componentsseparately may permit long-term storage without losing the activecomponents' functions. Alternatively, media can be provided pre-mixed.

The ingredients included in the kits can be supplied in containers ofany sort such that the life of the different ingredients are preservedand are not adsorbed or altered by the materials of the container. Forexample, sealed glass ampules may contain ingredients that have beenpackaged under a neutral, non-reacting gas, such as nitrogen. Ampulesmay consist of any suitable material, such as glass, organic polymers,such as polycarbonate, polystyrene, etc., ceramic, metal or any othermaterial typically employed to hold similar ingredients. Other examplesof suitable containers include simple bottles that may be fabricatedfrom similar substances as ampules, and envelopes, that may comprisefoil-lined interiors, such as aluminum or an alloy. Other containersinclude test tubes, vials, flasks, bottles, syringes, or the like.Containers may have a sterile access port, such as a bottle having astopper that can be pierced. Other containers may have two compartmentsthat are separated by a readily removable membrane that upon removalpermits the ingredients to be mixed. Removable membranes may be glass,plastic, rubber, etc.

As stated, kits can be supplied with instructional materials.Instructions may be printed on paper or other substrate, and/or may besupplied as an electronic-readable media, such as a floppy disc, CD-ROM,DVD-ROM, Zip disc, videotape, audiotape, etc. Detailed instructions maynot be physically associated with the kit; instead, a user may bedirected to an internet web site specified by the manufacturer ordistributor of the kit, or supplied as electronic mail.

One advantage of the disclosed embodiments is that the methods are morerobust than those previously used producing plants that do not requirespecial treatments required by those produced using methods of the priorart. For example, methods disclosed herein do not require the use ofseeds or inflorescence to start plants; do not require selection ofdiseased starting plants (such as those exhibiting symptoms of witchesbroom or little leaf disease); do not require use of somaticembryogenesis and do not utilize pseudospiklets. For successful growthfollowing tissue culture, the produced plants do not require wateringdirectly on the pot but remain robust with overhead watering and do notrequire multiple adjustments to light intensity or humidity conditionsprior to transfer to a greenhouse or other growing conditions. Theseimprovements over prior methods provide even additional advantagesrelated to the health of produced plants and efficiency of growth andprocessing.

Non-limiting embodiments encompassed by the present disclosure include(Stage 1, Stage 2, Stage 3, etc, media are defined elsewhere herein):

I. The following species: Arundinaria gigantea; Bambusa balcoa; Bambusavulgaris; Bambusa vulgaris ‘Vitatta’; Bambusa Oldhamii; Bambusa tulda;endrocalamus brandesii; Dendrocalamus asper; Dendrocalamus hamiltoni;Dendrocalamus giganteus; Dendrocalamus membranaceus; Dendrocalamusstrictus; Gigantochloa aspera; Gigantochloa scortechini; Guadua culeata;uadua aculeata ‘Nicaragua’; Guadua amplexifolia; Guadua angustifolia;Guadua angustofolia bi-color; Guadua paniculata; Melocanna bambusoides;eohouzeaua dullooa (Teinostachyum); Ochlandra travancorica;Phyllostachys edulis ‘Moso’; Phyllostachys nigra; Phyllostachys nigra‘Henon’; Schizostachyum lumampao;

II. Stage 1 media: Media b-12-c-v media or b-10-v media;

III. Stage 2 media: CW1-v media; CW2-v media; CW3-v media; CW4-v media;CW5-v media; or CW6-v media for 10-120 day cycles; and

IV. Stage 3 media: Br-2-v media; Ech-v media or Amel-v media.

More particularly, the following embodiments can be used (Stage 1, Stage2, Stage 3, etc, media are defined elsewhere herein):

Starting with a bamboo plant between the ages of 3 months and 3 years, anode from the cane with the lateral shoot just breaking the sheath canbe used as the explant. Each nodal section can be cut into 3-5millimeter sections with the shoot intact. The outer sheaths can bepeeled off and discarded and the remaining nodal section piece put intoa 10% bleach solution with a final concentration of 0.6% sodiumhydrochloride. The explant in bleach solution can be placed onto a LabRotators, Adjustable speed, Barnstead/Lab line orbital Shaker (modelnumber KS 260) shaker table for 1 hour at 6-9 revolutions per minute.The explants can then be put into a 1% bleach solution with a finalconcentration of 0.06% sodium hydrochloride, and be placed back onto theshaker table for 30 minutes. This 1% bleach solution step can then berepeated.

Individual explants can then be placed on a Stage 1 media (15-25 mL)within a tube and the tubes can be placed into a regulated clean growthchamber at a temperature of from 65° F.-70° F. and a full spectrum lightlevel of 200-500 foot candles. The initial Stage 1 media can be b-12c-ivat a pH of 5.7. The explants can then be transferred to fresh b-12c-ivmedia every 10-120 days (usually every 21 days), with contaminated tubesbeing discarded. Contaminated tubes can be identified by bacterialdiscoloration of the agar or by visible surface contamination. Theseexplants can stay on b-12c-iv media for 3-4 10-120 day cycles (usually21 day cycles). Explants can then be taken off the media after the thirdcycle if multiplication is occurring. If multiplication is not occurringor not occurring to a significant degree, explants can be left on themedia for a fourth cycle.

Live shoots can next be transferred to a Stage 2 media, such as b-9,CW1, CW2, CW3, CW4, CW5, CW6 or b-6 at a pH of 5.7. The cultures canstay on this Stage 2 media until the desired number of shoots isobtained by separation into new tubes and further expansion. Generally,the range of time includes 10-120 day cycles (usually 14-21 day cycles)between which the cultures are assigned to go through anothermultiplication round in Stage 2 media or transitioned to a Stage 3media, for example, b-10-iv or b-11-iv at a pH of 5.7 for furthermultiplication. One-ten shoots per tube can be obtained permultiplication cycle.

Following removal from the multiplication process, the shoots cantransferred to small tissue culturing boxes (known as “magenta boxes”)for 10-120 days (usually 14-21 days) containing a Stage 3 or Stage 4media, in this Example, BR-2 at a pH of 5.7 for 10-120 days (usually14-21 days) or Amel at a pH of 5.7 for 10-120 days (usually 14-21 days).

The following procedures may also be used (Stage 1, Stage 2, Stage 3,etc, media are defined elsewhere herein):

Starting with a bamboo plant between the ages of 3 months and 3 years, anode from the cane with the lateral shoot just breaking the sheath canbe used as the explant. Each nodal section can be cut into 3-5millimeter sections with the shoot intact. Some explants, includingexplants taken from canes 1 year or older can be pre-rinsed by shakingthem in a jar of 70% isopropyl alcohol for 3 seconds followed by rinsingthem under running tap water for 1 minute. Other explants are notpre-rinsed.

The outer sheaths can be peeled off and discarded and the remainingnodal section piece put into a 10% bleach solution. The explant inbleach solution can be placed onto a Lab Rotators, Adjustable speed,Barnstead/Lab line orbital Shaker (model number KS 260) shaker table for1 hour at 6-9 revolutions per minute. For some implants, including thosetaken from canes 1 year or older, this step can be modified by adding afew drops of Tween 20 to the 10% bleach solution and soaking theexplants for 45 minutes rather than 1 hour. The explants can then be putinto a 1% bleach solution, and placed back onto the shaker table for 30minutes. This 1% bleach solution step can then be repeated.

Individual explants can then be placed on a Stage 1 media (15-25 mL)within a tube and the tubes placed into a regulated clean growth chamberat a temperature of from 65° F.-70° F. and a full spectrum light levelof 200-500 foot candles. The Stage 1 media can be b-12c-iv at a pH of5.7. The explants can be transferred to fresh b-12c-iv media every10-120 days (usually every 21 days), with contaminated tubes beingdiscarded. These explants can stay on b-12c-iv media for 2 10-120 daycycles (usually 21 day cycles). Between cycles, excess sheaths can beremoved. At the time of transfer to the third cycle, explants can betransitioned to a Stage 2 media, in this Example, b-12-c supplementedwith 7 g/L carageenan rather than the 5.5 g/L provided above. Followingthe third cycle, explants can be cleaned. The explants can be kept onb-12-c supplemented with 7 g/L carageenan rather than the 5.5 g/Lprovided above for 10-120 day cycles (usually 21 day cycles) untilmultiple shoots are observed. Observation of multiple shoots can occurwithin 3-15 months.

Once the explant exhibits multiple shoots, it can be either maintainedon its Stage 2 media or transferred to a Stage 3 media. Non-limitingStage 3 media include, a b-9 media, a CW1 media, a CW2 media a CW3media, a CW4 media, a CW5 media, a CW6 media or a b-6 media at a pH of5.7. The cultures can stay on Stage 2 or Stage 3 media until the desirednumber of shoots is obtained by separation into new tubes and furtherexpansion. Generally, the range of time includes 10-120 day cycles(usually 21 day cycles) between which the cultures can be assigned to gothrough another multiplication round or transitioned to a Stage 3 orStage 4 media, such as a BR-2 media at a pH of 5.7 for 10-120 days(usually 21 days) in “magenta boxes” or a Amel media at a pH of 5.7 for10-120 days (usually 14-21 days).

In even more particular non-limiting embodiments, the following speciescan be micropropagated in the following media according to proceduresdescribed in the proceeding paragraphs [000137]-[000145], usually as aStage 2 media, at a pH of 5.5-5.7:

Arundinaria gigantea: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Bambusa balcoa: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Bambusa vulgaris: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v,

Bambusa vulgaris ‘Vitatta’: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Bambusa Oldhamii: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Bambusa tulda: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Dendrocalamus brandesii: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Dendrocalamus asper: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-V;

Dendrocalamus hamiltoni: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Dendrocalamus giganteus: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Dendrocalamus membranaceus: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Dendrocalamus strictus: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Gigantochloa aspera: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Gigantochloa scortechini: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Guadua culeata: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Guadua aculeata ‘Nicaragua’: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Guadua amplexifolia: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Guadua angustifolia: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Guadua angustofolia bi-color: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v orCW6-v;

Guadua paniculata: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Melocanna bambusoides: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Neohouzeaua dullooa (Teinostachyum): b-9-v, CW1-v, CW3-v, CW4-v, CW5-vor CW6-v;

Ochlandra travancorica: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Phyllostachys edulis ‘Moso’: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Phyllostachys nigra: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Phyllostachys nigra ‘Henan’: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v;

Schizostachyum lumampao: b-9-v, CW1-v, CW3-v, CW4-v, CW5-v or CW6-v.

As stated previously, there are many uses for bamboos produced accordingto the methods disclosed herein. In addition to or complementing thoseuses described elsewhere, non-limiting examples of uses and productsmade from bamboos produced according to the media, systems and methodsdisclosed herein include:

Exemplary Paper Types: Freesheet; Stock; Acid-free; A4; Board; Bond;Book; Bristol; Carbonless; Catalog; Coated; Cover; Dual-Purpose Bond;Duplex; English Finish; Equivalent; Fine; Free Sheet; Grain Long; GrainShort; Groundwood; Kraft; Lightweight; News Print; Publishing; Rag;Recycled; Tag; Uncoated; Virgin; Absorbent; Acid; Album; Albumin;Alkaline; Bank Note; Tissue; Toilet; Towels; Fluff; Card Stock; one-timecarbon (OTC); optical character recognition (OCR); Tissue Overlay; andNapkins.

Exemplary Pulp Types: Air-dried; Alpha; Bamboo; Bisulfate; Sulphite;Bleached; Chemical Cellulose (Dissolving); Fluff; Fodder; Free; FullyBleached; Hard; High Alpha Cellulose; Groundwood; Hot Groundwood; Jute;Knotter; Kraft; Long Fiber; Baled; Rolled; Market; Non-Wood; Board;Pressurized Groundwood; Rag; Recycled; Reinforcement; Secondary;Semi-alkaline; Semi-bleached; Semi-chemical; Short Fiber; Soda;Specialty; Sulfate; Thermochemical; Unbleached; Viscose; and Wood.

Board/Containers: Linerboard; Containerboard; Cardboard; old corrugatedcontainers (OCC); and Paperboard.

Wood: Structural Wood Panels (including Structural Plywood; OrientedStrand Board; Structural Composite Panels); Glued Laminated Timber;Structural Composite Lumber (including Laminated Veneer Lumber; ParallelStrand Lumber; Oriented Strand Lumber); Prefabricated Wood I-Joists;Floor Joints; Railroad Ties; Flooring; and Composites (including Auto;Aero; Musical).

Textiles: Feedstock; Filament Yarn; Knitted Fabric; Knitting; NarrowWidth Fabric; Non Woven Fabric; Spun Yarn; Woven Fabric; Viscose Rayon;Batting; Ginned Fiber; and Cloth.

Textile Products: Clothing; Towels; Sheets/Bedding; Pillows; Curtains

Food sources: Shoots; and any direct or bi-product for food consumptionby animals and humans.

Consumer Goods: Animal Feed; Carpeting; Light Bulbs; Household CleaningProducts; Chopsticks & Toothpicks; Cleaning Brooms; Bicycles; WheelChairs; Fishing rods; Beer; Liquor; Pharmaceuticals; Cosmetics;Soap/Shampoo; Kitchenware; Crafts; Furniture; Nutraceuticals; Papercups; Paper plates; and Diapers.

Energy & Bioenergy: Charcoal; Insulation; Feedstock; and Biomass.

The following non-limiting examples are provided. In all examples, timeon media that supports transition to ex vitro conditions can be in airpermeable or air impermeable containers. Described media are utilized intheir solid forms provided above unless otherwise noted as liquid.

EXAMPLES Example 1 Phyllostachys bissetti

Starting with a bamboo plant between the ages of 3 months and 3 years, anode from the cane with the lateral shoot just breaking the sheath wasused as the explant. Each nodal section was cut into 3-5 millimetersections with the shoot intact. The outer sheaths were peeled off anddiscarded and the remaining nodal section piece put into a 10% bleachsolution with a final concentration of 0.6% sodium hydrochloride. Theexplant in bleach solution was placed onto a Lab Rotators, Adjustablespeed, Barnstead/Lab line orbital Shaker (model number KS 260) shakertable for 1 hour at 6-9 revolutions per minute. The explants were thenput into a 1% bleach solution with a final concentration of 0.06% sodiumhydrochloride, and placed back onto the shaker table for 30 minutes.This 1% bleach solution step was then repeated.

Individual explants were then placed on a Stage 1 media (15-25 mL)within a tube and the tubes were placed into a regulated clean growthchamber at a temperature of from 65° F.-70° F. and a full spectrum lightlevel of 200-500 foot candles. The initial Stage 1 media in this Examplewas b-12c-iv at a pH of 5.7. The explants were transferred to freshb-12c-iv media every 10-120 days (usually every 21 days), withcontaminated tubes being discarded. Contaminated tubes were identifiedby bacterial discoloration of the agar or by visible surfacecontamination. These explants stayed on b-12c-iv media for 3-4 10-120day cycles (usually 21 day cycles). Explants were taken off the mediaafter the third cycle if multiplication was occurring. If multiplicationwas not occurring or not occurring to a significant degree, explantswere left on the media for a fourth cycle.

Live shoots were next transferred to a Stage 2 media, in this Example,b-9-iv at a pH of 5.7. The cultures stayed on b-9-iv media until thedesired number of shoots was obtained by separation into new tubes andfurther expansion. Generally, the range of time included 10-120 daycycles (usually 14-21 day cycles) between which the cultures wereassigned to go through another multiplication round in Stage 2 media ortransitioned to a Stage 3 media, in this Example, b-10-iv at a pH of 5.7for further multiplication. One-ten shoots per tube were obtained permultiplication cycle.

Following removal from the multiplication process, the shoots weretransferred to small tissue culturing boxes (known as “magenta boxes”)for 10-120 days (usually 14-21 days) containing a Stage 3 or Stage 4media, in this Example. BR-2-iv at a pH of 5.7 for 10-120 days (usually14-21 days).

Example 2 Fargesia denudata

In the example of Fargesia denudata, the explants were chosen anddisinfected as in Example 1.

The explants were then transferred into jars containing a Stage 1 media,in this Example, b-12c-iv (liquid; 30-40 mL) as described in Example 1but for the use of jars. Explants were taken off the media after thethird cycle if multiplication was occurring. If multiplication was notoccurring or not occurring to a significant degree, explants were lefton the media for a fourth cycle. Contaminated tubes were discarded.

The cultures were then transferred onto a Stage 2 media, in thisExample, b-11-iv (liquid) in jars on a rotating shelf that provides 6-9revolutions per minute. The cultures remained on b-11-iv media at a pHof 5.7 for 10-120 day cycles (usually 14 day cycles) until the desirednumber of shoots was obtained by separation into new jars and furtherexpansion. One-fifteen shoots per jar were obtained per multiplicationcycle. The shoots were then placed in a Stage 3 media, in this Example,Ech-iv at a pH of 6 for 10-120 days (usually 14-21 days).

Example 3 Pleioblastus fortunei

In the example of Pleioblastus fortunei, the explants were chosen anddisinfected as in Example 1. The explants were then transferred intotubes containing a Stage 1 media, in this Example, b-12c-iv also asdescribed in Example 1. Shoots were then transferred to a Stage 2 media,in this Example, b-9-iv in magenta boxes (40-50 mL). They remained onb-9-iv media for 10-120 day cycles (usually 14 day cycles) until thedesired number of shoots was obtained by separation into new boxes andfurther expansion. One-twenty shoots per box were obtained permultiplication cycle. The shoots were then placed on a Stage 3 media, inthis Example, BR-2-iv for 10-120 days (usually 14-21 days).

Example 4 Sasa Veitchii

In the example of Sasa Veitchii, the explants were chosen anddisinfected as in Example 1.

The explants were then transferred into tubes containing a Stage 1media, in this Example, b-12c-iv also as described in Example 1. Shootswere then transferred into a Stage 2 media, in this Example, b-1-iv at apH of 5.5 for 10-120 day cycles (usually 21 day cycles) until thedesired number of shoots was obtained by separation into new tubes andfurther expansion One-ten shoots per tube were obtained permultiplication cycle. The shoots were then placed in a Stage 3 media, inthis Example, Br-2-iv at a pH of 5.7 for 14-21 days.

Example 5 Pleioblastus viridistriatus and Thamnocalamus crassinodus

In the example of Pleioblastus viridistriatus and Thamnocalamuscrassinodus, the explants were chosen and disinfected as in Example 1.The explants were then transferred into tubes containing a Stage 1media, in this Example, b-12c-iv also as described in Example 1. Shootswere then transferred into a Stage 2 media, in this Example, b-4-iv at apH of 5.5 for 10-120 day cycles (usually 21 day cycles) until thedesired number of shoots was obtained by separation into new tubes andfurther expansion. One-ten shoots per tube were obtained permultiplication cycle. The shoots were then placed in a Stage 3 media, inthis Example, Br-2-iv at a pH of 5.7 for 10-120 days (usually 14-21days).

Example 6 Chusquea Culeo “Cana Prieta”

In the example of Chusquea Culeo “Cana Prieta”, the explants were chosenand disinfected as in Example 1. The explants were then transferred intotubes containing a Stage 1 media, in this Example, b-12c-iv media alsoas described in Example 1. Shoots were then transferred into a Stage 2media, in this Example, b-9-iv at a pH of 5.5 for 10-120 day cycles(usually 21 day cycles) until the desired number of shoots was obtainedby separation into new tubes and further expansion. One-ten shoots pertube were obtained per multiplication cycle. The shoots were then placedin a Stage 3 media, in this Example, Amel-iv at a pH of 5.7 for 10-120days (usually 14-21 days).

Example 7 Bambusa Old Hamii

In the example of Bambusa Old hamii, the explants were chosen anddisinfected as in Example 1. The explants were then transferred intoboxes containing a Stage 1 media, in this Example, b-10-iv (40-50 mL)also as described in Example 1 but for the change to boxes. Shoots weremaintained on b-10-iv media for 10-120 day cycles (usually 21 daycycles) until the desired number of shoots was obtained by separationinto new boxes and further expansion. One-twenty shoots per box wereobtained per multiplication cycle. The shoots were then placed in aStage 2 media, in this Example, Amel-iv at a pH of 5.7 for 10-120 days(usually 14-21 days).

Example 8 Phyllostachys Edulis “Moso”, Phyllostachys Atrovaginata &Dendrocalamus Asper

In the example of Phyllostachys Edulis “Moso”, PhyllostachysAtrovaginata & Dendrocalamus Asper, the explants were chosen anddisinfected as in Example 1. The explants were then transferred intotubes containing a Stage 1 media, in this Example, b-12c-iv also asdescribed in Example 1. Shoots were then transferred into a Stage 2media, in this Example, b-9-iv at a pH of 5.5 for 10-120 day cycles(usually 21 day cycles) until the desired number of shoots was obtainedby separation into new tubes and further expansion. A B-6 media at a pHof 5.5 can also be used. One-ten shoots per tube were obtained permultiplication cycle. The shoots were then placed in a Stage 3 media, inthis Example, Amel-iv at a pH of 5.7 for 10-120 days (usually 14-21days).

Example 9 Guadua Angustifolia

In the example of Guadua Angustofolia, the explants were chosen anddisinfected as in Example 1. The explants were then transferred intotubes containing a Stage 1 media, in this Example, b-12c-iv also asdescribed in Example 1. Shoots were then transferred into a Stage 2media, in this Example, b-10-iv at a pH of 5.5 for 10-120 day cycles(usually 21 day cycles) until the desired number of shoots was obtainedby separation into new tubes and further expansion. One-ten shoots pertube were obtained per multiplication cycle. The shoots were then placedin a Stage 3 media, in this Example, Amel-iv at a pH of 5.7 for 10-120days (usually 14-21 days).

Example 10 Phyllostachys bissetti—Alternate Procedure

Starting with a bamboo plant between the ages of 3 months and 3 years, anode from the cane with the lateral shoot just breaking the sheath wasused as the explant. Each nodal section was cut into 3-5 millimetersections with the shoot intact. Some explants, including explants takenfrom canes 1 year or older were pre-rinsed by shaking them in a jar of70% isopropyl alcohol for 3 seconds followed by rinsing them underrunning tap water for 1 minute. Other explants were not pre-rinsed.

The outer sheaths were peeled off and discarded and the remaining nodalsection piece put into a 10% bleach solution. The explant in bleachsolution was placed onto a Lab Rotators, Adjustable speed, Barnstead/Labline orbital Shaker (model number KS 260) shaker table for 1 hour at 6-9revolutions per minute. For some implants, including those taken fromcanes 1 year or older, this step was modified by adding a few drops ofTween 20 to the 10% bleach solution and soaking the explants for 45minutes rather than 1 hour. The explants were then put into a 1% bleachsolution, and placed back onto the shaker table for 30 minutes. This 1%bleach solution step was then repeated.

Individual explants were then placed on a Stage 1 media (15-25 mL)within a tube and the tubes were placed into a regulated clean growthchamber at a temperature of from 65° F.-70° F. and a full spectrum lightlevel of 200-500 foot candles. In this Example, the Stage 1 media wasb-12c-iv at a pH of 5.7. The explants were transferred to fresh b-12c-ivmedia every 10-120 days (usually every 21 days), with contaminated tubesbeing discarded. These explants stayed on b-12c-iv media for 2 10-120day cycles (usually 21 day cycles). Between cycles, excess sheaths wereremoved. At the time of transfer to the third cycle, explants weretransitioned to a Stage 2 media, in this Example, b-12-c supplementedwith 7 g/L carageenan rather than the 5.5 g/L provided above. Followingthe third cycle, explants were cleaned. The explants were kept on b-12-csupplemented with 7 g/L carageenan for 10-120 day cycles (usually 21 daycycles) until multiple shoots were observed. Observation of multipleshoots occurred within 3-15 months.

Once the explant exhibited multiple shoots, it was either maintained onits Stage 2 media or transferred to a Stage 3 media, in this Example,when used b-9-iv at a pH of 5.7. Alternatively to using one of the B-9media, a CW1 media at a pH of 5.7 can also be used. The cultures stayedon Stage 2 or Stage 3 media until the desired number of shoots wasobtained by separation into new tubes and further expansion. Generally,the range of time included 10-120 day cycles (usually 21 day cycles)between which the cultures were assigned to go through anothermultiplication round or were transitioned to a Stage 3 or Stage 4 media,in this Example, BR-2-iv at a pH of 5.7 for 10-120 days (usually 21days) in “magenta boxes”.

Example 11 Fargesia denudata—Alternate Procedure

In the example of Fargesia denudata, the explants were chosen anddisinfected as in Example 10. The explants were then transferred intojars containing a Stage 1 media, in this Example, b-12c-iv (liquid;30-40 mL) as described in Example 10 but for the use of jars. Theseexplants stayed on b-12c-iv media for 2 10-120 day cycles (usually 21day cycles). Between cycles, excess sheaths were removed. At the time oftransfer to the third cycle, explants were transitioned to a Stage 2media, in this Example, b-12-c supplemented with 7 g/L carageenan ratherthan the 5.5 g/L provided above. Following the third cycle, explantswere cleaned. The explants were kept on b-12-c supplemented with 7 g/Lcarageenan for 10-120 day cycles (usually 21 day cycles) until multipleshoots were observed. Observation of multiple shoots occurred within3-15 months.

Once the explant exhibited multiple shoots, it was either maintained onits Stage 2 media or transferred to a Stage 3 media, in this Example,b-11-iv (liquid) at a pH of 5.7 in jars on a rotating shelf thatprovides 6-9 revolutions per minute. The cultures remained on Stage 2 orStage 3 media for 10-120 day cycles (usually 14 day cycles) until thedesired number of shoots was obtained by separation into new jars andfurther expansion. One-fifteen shoots per jar were obtained permultiplication cycle. The shoots were then placed in a Stage 3 or Stage4 media, in this Example, Ech-iv at a pH of 6 for 10-120 days (usually21 days).

Example 12 Pleioblastus fortunei—Alternate Procedure

In the example of Pleioblastus fortunei, the explants were chosen anddisinfected as in Example 10. The explants were then transferred intotubes containing a Stage 1 media in this Example, b-12c-iv also asdescribed in Example 10. These explants stayed on b-12c-iv media for 210-120 day cycles (usually 21 day cycles). Between cycles, excesssheaths were removed. At the time of transfer to the third cycle,explants were transitioned to a Stage 2 media, in this Example, b-12-csupplemented with 7 g/L carageenan rather than the 5.5 g/L providedabove. Following the third cycle, explants were cleaned. The explantswere kept on b-12-c supplemented with 7 g/L carageenan for 10-120 daycycles (usually 21 day cycles) until multiple shoots were observed.Observation of multiple shoots occurred within 3-15 months.

Once the explant exhibited multiple shoots, it was either maintained onits Stage 2 media or transferred to a Stage 3 media, in this Exampleb-9-iv in magenta boxes (40-50 mL). (CW1 media can also be used). Theyremained on b-9-iv media for 10-120 day cycles (usually 14 day cycles)until the desired number of shoots was obtained by separation into newboxes and further expansion. One-twenty shoots per box were obtained permultiplication cycle. The shoots were then placed in a Stage 3 or Stage4 media, in this Example, BR-2-iv for 10-120 days (usually 14-21 days).

Example 13 Sasa Veitchii—Alternate Procedure

In the example of Sasa Veitchii, the explants were chosen anddisinfected as in Example 10.

The explants were then transferred into tubes containing a Stage 1media, in this Example, b-12c-iv also as described in Example 10. Theseexplants stayed on b-12c-iv media for 2 10-120 day cycles (usually 21day cycles). Between cycles, excess sheaths were removed. At the time oftransfer to the third cycle, explants were transitioned to a Stage 2media, in this Example, b-12-c supplemented with 7 g/L carageenan ratherthan the 5.5 g/L provided above. Following the third cycle, explantswere cleaned. The explants were kept on b-12-c supplemented with 7 g/Lcarageenan for 10-120 day cycles (usually 21 day cycles) until multipleshoots were observed. Observation of multiple shoots occurred within3-15 months.

Once the explant exhibited multiple shoots, it was either maintained onits Stage 2 media or transferred to a Stage 3 media, in this Exampleb-1-iv at a pH of 5.5 for 10-120 day cycles (usually 21 day cycles)until the desired number of shoots was obtained by separation into newtubes and further expansion One-ten shoots per tube were obtained permultiplication cycle. The shoots were then placed in a Stage 3 or Stage4 media, in this Example, Br-2-iv at a pH of 5.7 for 10-120 days(usually 14-21 days).

Example 14 Pleioblastus viridistriatus and Thamnocalamuscrassinodus—Alternate Procedure

In the example of Pleioblastus viridistriatus and Thamnocalamuscrassinodus, the explants were chosen and disinfected as in Example 10.The explants were then transferred into tubes containing a Stage 1media, in this Example, b-12c-iv also as described in Example 10. Theseexplants stayed on b-12c-iv media for 2 10-120 day cycles (usually 21day cycles). Between cycles, excess sheaths were removed. At the time oftransfer to the third cycle, explants were transitioned to a Stage 2media, in this Example, b-12-c supplemented with 7 g/L carageenan ratherthan the 5.5 g/L provided above. Following the third cycle, explantswere cleaned. The explants were kept on b-12-c supplemented with 7 g/Lcarageenan for 10-120 day cycles (usually 21 day cycles) until multipleshoots were observed. Observation of multiple shoots occurred within3-15 months.

Once the explant exhibited multiple shoots, it was either maintained onits Stage 2 media or transferred to a Stage 3 media, in this Exampleb-4-iv at a pH of 5.5 for 10-120 day cycles (usually 21 day cycles)until the desired number of shoots was obtained by separation into newtubes and further expansion. One-ten shoots per tube were obtained permultiplication cycle. The shoots were then placed in a Stage 3 or Stage4 media, in this Example, Br-2-iv at a pH of 5.7 for 10-120 days(usually 14-21 days).

Example 15 Chusquea Culeo “Cana Prieta”—Alternate Procedure

In the example of Chusquea Culeo “Cana Prieta”, the explants were chosenand disinfected as in Example 10. The explants were then transferredinto tubes containing a Stage 1 media, in this Example, b-12c-iv also asdescribed in Example 10. These explants stayed on b-12c-iv media for 210-120 day cycles (usually 21 day cycles). Between cycles, excesssheaths were removed. At the time of transfer to the third cycle,explants were transitioned to a Stage 2 media, in this Example, b-12-csupplemented with 7 g/L carageenan rather than the 5.5 g/L providedabove. Following the third cycle, explants were cleaned. The explantswere kept on b-12-c supplemented with 7 g/L carageenan for 10-120 daycycles (usually 21 day cycles) until multiple shoots were observed.Observation of multiple shoots occurred within 3-15 months.

Once the explant exhibited multiple shoots, it was either maintained onits Stage 2 media or transferred to a Stage 3 media, in this Exampleb-9-iv at a pH of 5.5 for 10-120 day cycles (usually 21 days) until thedesired number of shoots was obtained by separation into new tubes andfurther expansion. A B-6 media at a pH of 5.5 can also be used. One-tenshoots per tube were obtained per multiplication cycle. The shoots werethen placed in a Stage 3 or Stage 4 media, in this Example, Amel-ivmedia at a pH of 5.7 for 10-120 days (usually 14-21 days).

Example 16 Bambusa Old Hamii—Alternate Procedure

In the example of Bambusa Old hamii, the explants were chosen anddisinfected as in Example 10. The explants were then transferred intoboxes containing a Stage 1 media, in this Example, b-10-iv (40-50 mL)also as described in Example 10 but for the change to boxes. Theseexplants stayed on b-10-iv media for 2 10-120 day cycles (usually 21 daycycles). Between cycles, excess sheaths were removed. At the time oftransfer to the third cycle, explants were transitioned to a Stage 2media, in this Example, b-10-c supplemented with 7 g/L carageenan ratherthan the 5.5 g/L provided above. Following the third cycle, explantswere cleaned. The explants were kept on b-10-c supplemented with 7 g/Lcarageenan for 10-120 day cycles (usually 21 day cycles) until multipleshoots were observed. Observation of multiple shoots occurred within3-15 months. Cultures were maintained on Stage 2 media until the desirednumber of shoots was obtained. One-twenty shoots per box were obtainedper multiplication cycle. The shoots were then placed in a Stage 3media, in this Example, Amel-iv at a pH of 5.7 for 10-120 days (usually14-21 days).

Example 17 Phyllostachys Moso, Phyllostachys Atrovaginata &Dendrocalamus Asper—Alternate Procedure

In the example of Phyllostachys Moso, Phyllostachys Atrovaginata andDendrocalamus Asper, the explants were chosen and disinfected as inExample 10. The explants were then transferred into tubes containing aStage 1 media, in this Example, b-12c-iv also as described in Example10. These explants stayed on b-12c-iv media for 2 10-120 day cycles(usually 21 day cycles). Between cycles, excess sheaths were removed. Atthe time of transfer to the third cycle, explants were transitioned to aStage 2 media, in this Example, b-12-c supplemented with 7 g/Lcarageenan rather than the 5.5 g/L provided above. Following the thirdcycle, explants were cleaned. The explants were kept on b-12-csupplemented with 7 g/L carageenan for 10-120 day cycles (usually 21 daycycles) until multiple shoots were observed. Observation of multipleshoots occurred within 3-15 months.

Once the explant exhibited multiple shoots, it was either maintained onits Stage 2 media or transferred to a Stage 3 media, in this Exampleb-9-iv at a pH of 5.5 for 10-120 day cycles (usually 21 day cycles)until the desired number of shoots was obtained by separation into newtubes and further expansion. A B-6 media at a pH of 5.5 can also beused. One-ten shoots per tube were obtained per multiplication cycle.The shoots were then placed in a Stage 3 or Stage 4 media, in thisExample, Amel-iv at a pH of 5.7 for 10-120 days (usually 14-21 days).

Example 18 Guadua Angustifolia—Alternate Procedure

In the example of Guadua Angustofolia, the explants were chosen anddisinfected as in Example 10. The explants were then transferred intotubes containing a Stage 1 media, in this Example, b-12c-iv also asdescribed in Example 10. These explants stayed on b-12c-iv media for 210-120 day cycles (usually 21 day cycles). Between cycles, excesssheaths were removed. At the time of transfer to the third cycle,explants were transitioned to a Stage 2 media, in this Example, b-12-csupplemented with 7 g/L carageenan rather than the 5.5 g/L providedabove. Following the third cycle, explants were cleaned. The explantswere kept on b-12-c supplemented with 7 g/L carageenan for 10-120 daycycles (usually 21 day cycles) until multiple shoots were observed.Observation of multiple shoots occurred within 3-15 months.

Once the explant exhibited multiple shoots, it was either maintained onits Stage 2 media or transferred to a Stage 3 media, in this Exampleb-10-iv at a pH of 5.5 for 10-120 day cycles (usually 21 day cycles)until the desired number of shoots was obtained by separation into newtubes and further expansion. One-ten shoots per tube were obtained permultiplication cycle. The shoots were then placed in a Stage 3 or Stage4 media, in this Example, Amel-iv at a pH of 5.7 for 10-120 days(usually 14-21 days).

As will be understood by one of ordinary skill from the providedexamples, the tissue culturing method for individual species includesslight variations in media, timing and growth conditions. Thesevariations for individual species require optimization based on factorsincluding location, desired outcome, starting material, etc.

For each of the species provided in the examples listed above, inparticular embodiments, each can be initiated and/or multiplied in b-9-imedia, b-9-ii media, b-9-iii media, b-9-iv media, b-9-v media, CW2-imedia, CW2-ii media, CW2-iii media, CW2-iv media, CW2-v media, b-10-imedia, b-10-ii media, b-10-iii media, b-10-iv media, b-10-v media,b-11-i media, b-11-ii media, b-11-iii media, b-11-iv media, b-11-vmedia, b-12c-i media, b-12c-ii media, b-12c-iii media, b-12c-iv media,b-12c-v media, b-1-i media, b-1-ii media, b-1-iii media, b-1-iv media,b-1-v media, b-4-i media, b-4-ii media, b-4-iii media, b-4-iv media,b-4-v media, b-6-i media, b-6-ii media, b-6-iii media, b-6-iv media,b-6-v media, CW1-i media, CW1-ii media, CW1-iii media, CW1-iv media,CW1-v media, CW3-i media, CW3-ii media, CW3-iii media, CW3-iv media,CW3-v media, CW4-i media, CW4-ii media, CW4-iii media, CW4-iv media,CW4-v media, CW5-i media, CW5-ii media, CW5-iii media, CW5-iv media,CW5-v media, CW6-i media, CW6-ii media, CW6-iii media, CW6-iv mediaand/or CW6-v media.

As used herein “in” and “on” are interchangeable in the context ofplacing explants, shoots or plantlets within a tube, jar, box or jugcontaining a media.

Unless otherwise indicated, all numbers expressing quantities ofingredients, properties such as molecular weight, reaction conditions,and so forth used in the specification and claims are to be understoodas being modified in all instances by the term “about.” Accordingly,unless indicated to the contrary, the numerical parameters set forth inthe specification and attached claims are approximations that may varydepending upon the desired properties sought to be obtained by thepresent invention. At the very least, and not as an attempt to limit theapplication of the doctrine of equivalents to the scope of the claims,each numerical parameter should at least be construed in light of thenumber of reported significant digits and by applying ordinary roundingtechniques.

Notwithstanding that the numerical ranges and parameters setting forththe broad scope of the invention are approximations, the numericalvalues set forth in the specific examples are reported as precisely aspossible. Any numerical value, however, inherently contains certainerrors necessarily resulting from the standard deviation found in theirrespective testing measurements.

The terms “a,” “an,” “the” and similar referents used in the context ofdescribing the invention (especially in the context of the followingclaims) are to be construed to cover both the singular and the plural,unless otherwise indicated herein or clearly contradicted by context.Recitation of ranges of values herein is merely intended to serve as ashorthand method of referring individually to each separate valuefalling within the range. Unless otherwise indicated herein, eachindividual value is incorporated into the specification as if it wereindividually recited herein. All methods described herein can beperformed in any suitable order unless otherwise indicated herein orotherwise clearly contradicted by context. The use of any and allexamples, or exemplary language (e.g., “such as”) provided herein isintended merely to better illuminate the invention and does not pose alimitation on the scope of the invention otherwise claimed. No languagein the specification should be construed as indicating any non-claimedelement essential to the practice of the invention.

Groupings of alternative elements or embodiments of the inventiondisclosed herein are not to be construed as limitations. Each groupmember may be referred to and claimed individually or in any combinationwith other members of the group or other elements found herein. It isanticipated that one or more members of a group may be included in, ordeleted from, a group for reasons of convenience and/or patentability.When any such inclusion or deletion occurs, the specification is deemedto contain the group as modified thus fulfilling the written descriptionof all Markush groups used in the appended claims.

Certain embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention. Ofcourse, variations on these described embodiments will become apparentto those of ordinary skill in the art upon reading the foregoingdescription. The inventor expects skilled artisans to employ suchvariations as appropriate, and the inventors intend for the invention tobe practiced otherwise than specifically described herein. Accordingly,this invention includes all modifications and equivalents of the subjectmatter recited in the claims appended hereto as permitted by applicablelaw. Moreover, any combination of the above-described elements in allpossible variations thereof is encompassed by the invention unlessotherwise indicated herein or otherwise clearly contradicted by context.

Specific embodiments disclosed herein may be further limited in theclaims using consisting of or and consisting essentially of language.When used in the claims, whether as filed or added per amendment, thetransition term “consisting of” excludes any element, step, oringredient not specified in the claims. The transition term “consistingessentially of” limits the scope of a claim to the specified materialsor steps and those that do not materially affect the basic and novelcharacteristic(s). Embodiments of the invention so claimed areinherently or expressly described and enabled herein.

In closing, it is to be understood that the embodiments of the inventiondisclosed herein are illustrative of the principles of the presentinvention. Other modifications that may be employed are within the scopeof the invention. Thus, by way of example, but not of limitation,alternative configurations of the present invention may be utilized inaccordance with the teachings herein. Accordingly, the present inventionis not limited to that precisely as shown and described.

1.-19. (canceled)
 20. A method of micropropagating bamboo comprisinginitiating a shoot in vitro from a bamboo explant on a first media, andmultiplying the shoot initiated from the explant in vitro on a secondmedia, wherein the first media and/or the second media comprises (1)meta-topolin (mT) or analog thereof, (2) thidiazuron (TDZ) or analogthereof, (3) benzylaminopurine (BAP), and (4) β-naphthoxyacetic acid(NAA).
 21. The method of claim 20, wherein the bamboo explant is a nodalsection comprising a lateral shoot just breaking the sheath at the nodalsection.
 22. The method of claim 20, wherein the bamboo explant is froma bamboo plant ranging in age from about 3 months to about 3 years. 23.The method of claim 20, wherein at least three in vitro shoots areobtained from the explant following multiplication.
 24. The method ofclaim 23, wherein the method further comprises separating andindividually transferring the at least three in vitro shoots to freshfirst media or fresh second media.
 25. The method of claim 20, whereinthe method further comprises two or more cycles of growing the in vitroshoots on fresh first media and then on fresh second media.
 26. Themethod of claim 20, wherein the bamboo is Phyllostachys edulis.
 27. Themethod of claim 20, wherein the bamboo is Phyllostachys bissetti;Fargesia denudata; Pleioblastus fortunei; Sasa Veitchii; Pleioblastusviridistriatus; Thamnocalamus crassinodus; Chusquea Culeo “Cana Prieta”;Bambusa Old Hamii; Phyllostachys Atrovaginata; Dendrocalamus Asper;Guadua Angustifolia, Arundinaria gigante; Arundinaria gigantea ‘Macon’;Bambusa balcooa; Bambusa bambos; Bambusa lako; Bambusa stenostachya;Bambusa tulda; Bambusa vulgaris; Bambusa vulgaris ‘Vittata’; Chusqueaandina; Chusquea gigantea; Dendrocalamus brandisii; Dendrocalamusgiganteus; Dendrocalamus hamiltonii; Dendrocalamus latiflorus;Dendrocalamus membranaceus; Dendrocalamus strictus; Fargesia murieliae;Fargesia nitida; Fargesia nitida ‘Jiuzhaigou’; Fargesia robusta;Fargesia sp. ‘Scrabrida’; Gigantochloa apus; Gigantochloapseudoarundinacea; Guadua amplexifolia; Guadua chacoensis; Ochlandrastridula; Ochlandra travancorica; Phyllostachys bambusoides;Phyllostachys dulcis; Phyllostachys nigra; Phyllostachys nigra ‘Bory’;Phyllostachys nigra ‘Henon’; Phyllostachys nuda; Phyllostachysrubromarginata; Phyllostachys vivax; Pseudosasa japonica; orSemiarundinaria fastuosa.
 28. The method of claim 20, wherein the methodfurther comprises transferring the multiplied in vitro shoots to a thirdmedia that supports transition to ex vitro conditions.
 29. A method ofmicropropagating bamboo comprising initiating a shoot in vitro from abamboo explant on a first media comprising (1) meta-topolin (mT) oranalog thereof, (2) thidiazuron (TDZ) or analog thereof, (3)benzylaminopurine (BAP), and (4) β-naphthoxyacetic acid (NAA); andmultiplying the in vitro shoot on a second media comprising (1)meta-topolin (mT) or analog thereof, (2) benzylaminopurine (BAP), and(3) β-naphthoxyacetic acid (NAA).
 30. The method of claim 29, whereinthe bamboo explant is a nodal section comprising a lateral shoot justbreaking the sheath at the nodal section.
 31. The method of claim 29,wherein the bamboo explant is from a bamboo plant ranging in age fromabout 3 months to about 3 years.
 32. The method of claim 29, wherein atleast three in vitro shoots are obtained from the explant followingmultiplication.
 33. The method of claim 32, wherein the method furthercomprises separating and individually transferring the at least three invitro shoots to fresh first media or fresh second media.
 34. The methodof claim 29, wherein the method further comprises two or more cycles ofgrowing the in vitro shoots on fresh first media and then on freshsecond media.
 35. The method of claim 29, wherein the bamboo isPhyllostachys edulis.
 36. The method of claim 29, wherein the bamboo isPhyllostachys bissetti; Fargesia denudata; Pleioblastus fortunei; SasaVeitchii; Pleioblastus viridistriatus; Thamnocalamus crassinodus;Chusquea Culeo “Cana Prieta”; Bambusa Old Hamii; PhyllostachysAtrovaginata; Dendrocalamus Asper; Guadua Angustifolia; Arundinariagigante; Arundinaria gigantea ‘Macon’; Bambusa balcooa; Bambusa bambos;Bambusa lako; Bambusa stenostachya; Bambusa tulda; Bambusa vulgaris;Bambusa vulgaris ‘Vittata’; Chusquea andina; Chusquea gigantea;Dendrocalamus brandisii; Dendrocalamus giganteus; Dendrocalamushamiltonii; Dendrocalamus latiflorus; Dendrocalamus membranaceus;Dendrocalamus strictus; Fargesia murieliae; Fargesia nitida; Fargesianitida ‘Jiuzhaigou’; Fargesia robusta; Fargesia sp. ‘Scrabrida’;Gigantochloa apus; Gigantochloa pseudoarundinacea; Guadua amplexifolia;Guadua chacoensis; Ochlandra stridula; Ochlandra travancorica;Phyllostachys bambusoides; Phyllostachys dulcis; Phyllostachys nigra;Phyllostachys nigra ‘Bory’; Phyllostachys nigra ‘Henon’; Phyllostachysnuda; Phyllostachys rubromarginata; Phyllostachys vivax; Pseudosasajaponica; or Semiarundinaria fastuosa.
 37. The method of claim 29,wherein the method further comprises transferring the multiplied invitro shoots to a third media that supports transition to ex vitroconditions.
 38. A method of micropropagating bamboo in vitro comprisingusing at least one media comprising (1) meta-topolin (mT) or analogthereof, (2) thidiazuron (TDZ) or analog thereof, (3) benzylaminopurine(BAP), and (4) β-naphthoxyacetic acid (NAA).
 39. The method of claim 38,wherein the bamboo is Phyllostachys edulis.